Regulation of the Expression of DAPK1 by SUMO Pathway
Identifieur interne : 000426 ( Pmc/Curation ); précédent : 000425; suivant : 000427Regulation of the Expression of DAPK1 by SUMO Pathway
Auteurs : Qingshui Wang ; Xiuli Zhang ; Ling Chen ; Shuyun Weng ; Yun Xia ; Yan Ye ; Ke Li ; Ziqiang Liao ; Pengchen Chen ; Khaldoon Alsamman ; Chen Meng ; Craig Stevens ; Ted R. Hupp ; Yao LinSource :
- Biomolecules [ 2218-273X ] ; 2019.
Abstract
Death Associated Protein Kinase 1 (DAPK1) is an important signaling kinase mediating the biological effect of multiple natural biomolecules such as IFN-γ, TNF-α, curcumin, etc. DAPK1 is degraded through both ubiquitin-proteasomal and lysosomal degradation pathways. To investigate the crosstalk between these two DAPK1 degradation pathways, we carried out a screen using a set of ubiquitin E2 siRNAs at the presence of Tuberous Sclerous 2 (TSC2) and identified that the small ubiquitin-like molecule (SUMO) pathway is able to regulate the protein levels of DAPK1. Inhibition of the SUMO pathway enhanced DAPK1 protein levels and the minimum domain of DAPK1 protein required for this regulation is the kinase domain, suggesting that the SUMO pathway regulates DAPK1 protein levels independent of TSC2. Suppression of the SUMO pathway did not enhance DAPK1 protein stability. In addition, mutation of the potential SUMO conjugation sites on DAPK1 kinase domain did not alter its protein stability or response to SUMO pathway inhibition. These data suggested that the SUMO pathway does not regulate DAPK1 protein degradation. The exact molecular mechanism underlying this regulation is yet to be discovered.
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DOI: 10.3390/biom9040151
PubMed: 30999631
PubMed Central: 6523460
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Regulation of the Expression of DAPK1 by SUMO Pathway</title><author><name sortKey="Wang, Qingshui" sort="Wang, Qingshui" uniqKey="Wang Q" first="Qingshui" last="Wang">Qingshui Wang</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Zhang, Xiuli" sort="Zhang, Xiuli" uniqKey="Zhang X" first="Xiuli" last="Zhang">Xiuli Zhang</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Chen, Ling" sort="Chen, Ling" uniqKey="Chen L" first="Ling" last="Chen">Ling Chen</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Weng, Shuyun" sort="Weng, Shuyun" uniqKey="Weng S" first="Shuyun" last="Weng">Shuyun Weng</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Xia, Yun" sort="Xia, Yun" uniqKey="Xia Y" first="Yun" last="Xia">Yun Xia</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Ye, Yan" sort="Ye, Yan" uniqKey="Ye Y" first="Yan" last="Ye">Yan Ye</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Li, Ke" sort="Li, Ke" uniqKey="Li K" first="Ke" last="Li">Ke Li</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Liao, Ziqiang" sort="Liao, Ziqiang" uniqKey="Liao Z" first="Ziqiang" last="Liao">Ziqiang Liao</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Chen, Pengchen" sort="Chen, Pengchen" uniqKey="Chen P" first="Pengchen" last="Chen">Pengchen Chen</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Alsamman, Khaldoon" sort="Alsamman, Khaldoon" uniqKey="Alsamman K" first="Khaldoon" last="Alsamman">Khaldoon Alsamman</name><affiliation><nlm:aff id="af2-biomolecules-09-00151">Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Imam Abdulrahman bin Faisal University, Dammam 34212, Saudi Arabia;<email>kmalsamman@iau.edu.sa</email></nlm:aff></affiliation></author><author><name sortKey="Meng, Chen" sort="Meng, Chen" uniqKey="Meng C" first="Chen" last="Meng">Chen Meng</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Stevens, Craig" sort="Stevens, Craig" uniqKey="Stevens C" first="Craig" last="Stevens">Craig Stevens</name><affiliation><nlm:aff id="af3-biomolecules-09-00151">School of Applied Sciences, Edinburgh Napier University, Edinburgh EH11 4BN, UK;<email>C.Stevens@napier.ac.uk</email></nlm:aff></affiliation></author><author><name sortKey="Hupp, Ted R" sort="Hupp, Ted R" uniqKey="Hupp T" first="Ted R." last="Hupp">Ted R. Hupp</name><affiliation><nlm:aff id="af4-biomolecules-09-00151">Institute of Genetics and Molecular Medicine, Cell Signaling Unit, CRUK p53 Transduction Group, University of Edinburgh, EH4 2XR EH4 2XR, UK;<email>ted.hupp@ed.ac.uk</email></nlm:aff></affiliation></author><author><name sortKey="Lin, Yao" sort="Lin, Yao" uniqKey="Lin Y" first="Yao" last="Lin">Yao Lin</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">30999631</idno><idno type="pmc">6523460</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6523460</idno><idno type="RBID">PMC:6523460</idno><idno type="doi">10.3390/biom9040151</idno><date when="2019">2019</date><idno type="wicri:Area/Pmc/Corpus">000426</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000426</idno><idno type="wicri:Area/Pmc/Curation">000426</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000426</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Regulation of the Expression of DAPK1 by SUMO Pathway</title><author><name sortKey="Wang, Qingshui" sort="Wang, Qingshui" uniqKey="Wang Q" first="Qingshui" last="Wang">Qingshui Wang</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Zhang, Xiuli" sort="Zhang, Xiuli" uniqKey="Zhang X" first="Xiuli" last="Zhang">Xiuli Zhang</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Chen, Ling" sort="Chen, Ling" uniqKey="Chen L" first="Ling" last="Chen">Ling Chen</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Weng, Shuyun" sort="Weng, Shuyun" uniqKey="Weng S" first="Shuyun" last="Weng">Shuyun Weng</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Xia, Yun" sort="Xia, Yun" uniqKey="Xia Y" first="Yun" last="Xia">Yun Xia</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Ye, Yan" sort="Ye, Yan" uniqKey="Ye Y" first="Yan" last="Ye">Yan Ye</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Li, Ke" sort="Li, Ke" uniqKey="Li K" first="Ke" last="Li">Ke Li</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Liao, Ziqiang" sort="Liao, Ziqiang" uniqKey="Liao Z" first="Ziqiang" last="Liao">Ziqiang Liao</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Chen, Pengchen" sort="Chen, Pengchen" uniqKey="Chen P" first="Pengchen" last="Chen">Pengchen Chen</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Alsamman, Khaldoon" sort="Alsamman, Khaldoon" uniqKey="Alsamman K" first="Khaldoon" last="Alsamman">Khaldoon Alsamman</name><affiliation><nlm:aff id="af2-biomolecules-09-00151">Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Imam Abdulrahman bin Faisal University, Dammam 34212, Saudi Arabia;<email>kmalsamman@iau.edu.sa</email></nlm:aff></affiliation></author><author><name sortKey="Meng, Chen" sort="Meng, Chen" uniqKey="Meng C" first="Chen" last="Meng">Chen Meng</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author><author><name sortKey="Stevens, Craig" sort="Stevens, Craig" uniqKey="Stevens C" first="Craig" last="Stevens">Craig Stevens</name><affiliation><nlm:aff id="af3-biomolecules-09-00151">School of Applied Sciences, Edinburgh Napier University, Edinburgh EH11 4BN, UK;<email>C.Stevens@napier.ac.uk</email></nlm:aff></affiliation></author><author><name sortKey="Hupp, Ted R" sort="Hupp, Ted R" uniqKey="Hupp T" first="Ted R." last="Hupp">Ted R. Hupp</name><affiliation><nlm:aff id="af4-biomolecules-09-00151">Institute of Genetics and Molecular Medicine, Cell Signaling Unit, CRUK p53 Transduction Group, University of Edinburgh, EH4 2XR EH4 2XR, UK;<email>ted.hupp@ed.ac.uk</email></nlm:aff></affiliation></author><author><name sortKey="Lin, Yao" sort="Lin, Yao" uniqKey="Lin Y" first="Yao" last="Lin">Yao Lin</name><affiliation><nlm:aff id="af1-biomolecules-09-00151">Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</nlm:aff></affiliation></author></analytic><series><title level="j">Biomolecules</title><idno type="eISSN">2218-273X</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Death Associated Protein Kinase 1 (DAPK1) is an important signaling kinase mediating the biological effect of multiple natural biomolecules such as IFN-γ, TNF-α, curcumin, etc. DAPK1 is degraded through both ubiquitin-proteasomal and lysosomal degradation pathways. To investigate the crosstalk between these two DAPK1 degradation pathways, we carried out a screen using a set of ubiquitin E2 siRNAs at the presence of Tuberous Sclerous 2 (TSC2) and identified that the small ubiquitin-like molecule (SUMO) pathway is able to regulate the protein levels of DAPK1. Inhibition of the SUMO pathway enhanced DAPK1 protein levels and the minimum domain of DAPK1 protein required for this regulation is the kinase domain, suggesting that the SUMO pathway regulates DAPK1 protein levels independent of TSC2. Suppression of the SUMO pathway did not enhance DAPK1 protein stability. In addition, mutation of the potential SUMO conjugation sites on DAPK1 kinase domain did not alter its protein stability or response to SUMO pathway inhibition. These data suggested that the SUMO pathway does not regulate DAPK1 protein degradation. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Biomolecules</journal-id><journal-id journal-id-type="iso-abbrev">Biomolecules</journal-id><journal-id journal-id-type="publisher-id">biomolecules</journal-id><journal-title-group><journal-title>Biomolecules</journal-title></journal-title-group><issn pub-type="epub">2218-273X</issn><publisher><publisher-name>MDPI</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">30999631</article-id><article-id pub-id-type="pmc">6523460</article-id><article-id pub-id-type="doi">10.3390/biom9040151</article-id><article-id pub-id-type="publisher-id">biomolecules-09-00151</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Regulation of the Expression of DAPK1 by SUMO Pathway</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Qingshui</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref><xref ref-type="author-notes" rid="fn1-biomolecules-09-00151">†</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Xiuli</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref><xref ref-type="author-notes" rid="fn1-biomolecules-09-00151">†</xref></contrib><contrib contrib-type="author"><name><surname>Chen</surname><given-names>Ling</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref><xref ref-type="author-notes" rid="fn1-biomolecules-09-00151">†</xref></contrib><contrib contrib-type="author"><name><surname>Weng</surname><given-names>Shuyun</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><name><surname>Xia</surname><given-names>Yun</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><name><surname>Ye</surname><given-names>Yan</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Ke</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><name><surname>Liao</surname><given-names>Ziqiang</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><name><surname>Chen</surname><given-names>Pengchen</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid" authenticated="true">https://orcid.org/0000-0003-0068-5516</contrib-id><name><surname>Alsamman</surname><given-names>Khaldoon</given-names></name><xref ref-type="aff" rid="af2-biomolecules-09-00151">2</xref></contrib><contrib contrib-type="author"><name><surname>Meng</surname><given-names>Chen</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref></contrib><contrib contrib-type="author"><name><surname>Stevens</surname><given-names>Craig</given-names></name><xref ref-type="aff" rid="af3-biomolecules-09-00151">3</xref></contrib><contrib contrib-type="author"><name><surname>Hupp</surname><given-names>Ted R.</given-names></name><xref ref-type="aff" rid="af4-biomolecules-09-00151">4</xref></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid" authenticated="true">https://orcid.org/0000-0002-0493-0155</contrib-id><name><surname>Lin</surname><given-names>Yao</given-names></name><xref ref-type="aff" rid="af1-biomolecules-09-00151">1</xref><xref rid="c1-biomolecules-09-00151" ref-type="corresp">*</xref></contrib></contrib-group><aff id="af1-biomolecules-09-00151"><label>1</label>Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China;<email>wangqingshui@fjnu.edu.cn</email> (Q.W.);<email>15980239296@163.com</email> (X.Z.);<email>chenling654321@163.com</email> (L.C.);<email>wsy09080700@163.com</email> (S.W.);<email>xiayunnyyl@163.com</email> (Y.X.);<email>m18759141945@163.com</email> (Y.Y.);<email>13107673087@163.com</email> (K.L.);<email>liaoziqiangcontact@163.com</email> (Z.L.);<email>yb77620@umac.mo</email> (P.C.);<email>mmenger@126.com</email> (C.M.)</aff><aff id="af2-biomolecules-09-00151"><label>2</label>Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Imam Abdulrahman bin Faisal University, Dammam 34212, Saudi Arabia;<email>kmalsamman@iau.edu.sa</email></aff><aff id="af3-biomolecules-09-00151"><label>3</label>School of Applied Sciences, Edinburgh Napier University, Edinburgh EH11 4BN, UK;<email>C.Stevens@napier.ac.uk</email></aff><aff id="af4-biomolecules-09-00151"><label>4</label>Institute of Genetics and Molecular Medicine, Cell Signaling Unit, CRUK p53 Transduction Group, University of Edinburgh, EH4 2XR EH4 2XR, UK;<email>ted.hupp@ed.ac.uk</email></aff><author-notes><corresp id="c1-biomolecules-09-00151"><label>*</label>Correspondence: <email>yaolin@fjnu.edu.cn</email>; Tel.: +86-(0)591-22860592</corresp><fn id="fn1-biomolecules-09-00151"><label>†</label><p>These authors contributed equally to this work.</p></fn></author-notes><pub-date pub-type="epub"><day>17</day><month>4</month><year>2019</year></pub-date><pub-date pub-type="collection"><month>4</month><year>2019</year></pub-date><volume>9</volume><issue>4</issue><elocation-id>151</elocation-id><history><date date-type="received"><day>15</day><month>3</month><year>2019</year></date><date date-type="accepted"><day>15</day><month>4</month><year>2019</year></date></history><permissions><copyright-statement>© 2019 by the authors.</copyright-statement><copyright-year>2019</copyright-year><license license-type="open-access"><license-p>Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>).</license-p></license></permissions><abstract><p>Death Associated Protein Kinase 1 (DAPK1) is an important signaling kinase mediating the biological effect of multiple natural biomolecules such as IFN-γ, TNF-α, curcumin, etc. DAPK1 is degraded through both ubiquitin-proteasomal and lysosomal degradation pathways. To investigate the crosstalk between these two DAPK1 degradation pathways, we carried out a screen using a set of ubiquitin E2 siRNAs at the presence of Tuberous Sclerous 2 (TSC2) and identified that the small ubiquitin-like molecule (SUMO) pathway is able to regulate the protein levels of DAPK1. Inhibition of the SUMO pathway enhanced DAPK1 protein levels and the minimum domain of DAPK1 protein required for this regulation is the kinase domain, suggesting that the SUMO pathway regulates DAPK1 protein levels independent of TSC2. Suppression of the SUMO pathway did not enhance DAPK1 protein stability. In addition, mutation of the potential SUMO conjugation sites on DAPK1 kinase domain did not alter its protein stability or response to SUMO pathway inhibition. These data suggested that the SUMO pathway does not regulate DAPK1 protein degradation. The exact molecular mechanism underlying this regulation is yet to be discovered.</p></abstract><kwd-group><kwd>DAPK1</kwd><kwd>SUMO</kwd><kwd>SENP</kwd><kwd>protein degradation</kwd><kwd>post-translational modification</kwd></kwd-group></article-meta></front><floats-group><fig id="biomolecules-09-00151-f001" orientation="portrait" position="float"><label>Figure 1</label><caption><p>Inhibition of SUMO pathway partially restored DAPK1 protein levels at the presence of TSC2. (<bold>A</bold>) HEK293T was transfected with HA-DAPK1, Flag-TSC2, and different E2 siRNAs. (<bold>B</bold>,<bold>C</bold>) HEK293T cells were transfected with HA-DAPK1, LacZ, TSC2 and four different SENPs as indicated. Cell lysates were extracted 48 h post-transfection and immunoblotted with respective antibodies. The intensity of the bands was quantified using LICOR Odyssey software and represented by bar graphs. The experiments were repeated three times (n = 3), and representative images are presented. The representative western blot images are from different gels and each lane was loaded with the same amount of proteins. Data of triplicate assays are presented as mean ± S.D. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001; NS, no significance. “+” indicated that the plasmid was transfected, “−” indicated that the plasmid was not transfected.</p></caption><graphic xlink:href="biomolecules-09-00151-g001"></graphic></fig><fig id="biomolecules-09-00151-f002" orientation="portrait" position="float"><label>Figure 2</label><caption><p>The SUMO pathway regulated the protein levels of DAPK1. HEK293T were transfected with (<bold>A</bold>) HA-DAPK1, LacZ, V5-UBC9, His-SUMO-1, His-SUMO-2 or His-SUMO-3, or (<bold>B</bold>) HA-DAPK1, LacZ, V5-UBC9, His-SUMO-1, His-SUMO-2 or His-SUMO-3, or (<bold>C</bold>) HA-DAPK1, LacZ and six different SENPs, or (<bold>E</bold>) control and UBC9 siRNA as indicated. HCT116 were transfected with (<bold>D</bold>) HA-DAPK1, LacZ and six different SENPs as indicated. Cell lysates were extracted 48 h post-transfection and immunoblotted with respective antibodies. The intensity of the bands was quantified using LICOR Odyssey software and represented by bar graphs. The experiments were repeated three times (n = 3) and representative images are presented. The representative western blot images are from different gels and each lane was loaded with the same amount of proteins. Data of triplicate assays are presented as mean ± S.D. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001; NS, no significance. “+” indicated that the plasmid was transfected, “−” indicated that the plasmid was not transfected.</p></caption><graphic xlink:href="biomolecules-09-00151-g002"></graphic></fig><fig id="biomolecules-09-00151-f003" orientation="portrait" position="float"><label>Figure 3</label><caption><p>SENPs up-regulated DAPK1 protein levels via the 1-364 kinase domain. (<bold>A</bold>) A diagram illustrating the panel of DAPK1 deletion mutants. (<bold>B–D</bold>) The DAPK1 deletion mutants were co-transfected with LacZ and either Flag-SENP1 (<bold>B</bold>), Flag-SENP2 (<bold>C</bold>) or Flag-SENP6 (<bold>D</bold>), as indicated. (<bold>E</bold>) HA-DAPK (275–1430) mutant was co-transfected with LacZ and either Flag-SENP1, Flag-SENP2 or Flag-SENP6. Cell lysates were extracted 48 h post-transfection and immunoblotted with respective antibodies. The intensity of the bands was quantified using LICOR Odyssey software and represented by bar graphs. The experiments were repeated three times (n = 3) and representative images are presented. The representative western blot images are from different gels and each lane was loaded with the same amount of proteins. Data of triplicate assays are presented as mean ± S.D. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001; NS, no significance. “+” indicated that the plasmid was transfected, “−” indicated that the plasmid was not transfected.</p></caption><graphic xlink:href="biomolecules-09-00151-g003"></graphic></fig><fig id="biomolecules-09-00151-f004" orientation="portrait" position="float"><label>Figure 4</label><caption><p>SUMO pathway did not regulate the protein degradation of HA-DAPK1 (1–364). (<bold>A</bold>) HEK293T cells or (<bold>B</bold>) HCT116 cells transfected with DAPK1 (1–364) and LacZ were exposed to MG132 (10 μM, 6 h) or leupeptin (200 μM), Est (10 μg/mL) and chloroquine (100 μM) for 24 h as indicated. (<bold>C</bold>) HEK293T transfected with LacZ, DAPK1(1–364) were exposed to 20 μg/mL CHX for 0–8 h as indicated. (<bold>D</bold>) HEK293T transfected with HA-DAPK1 (1–364) and LacZ were exposed to 10 μM MG132 and 20 μg/mL CHX for 0–8 h as indicated. (<bold>E</bold>) HEK293T transfected with LacZ, HA-DAPK1 (1–364) and Flag-SENP6 were exposed to 20 μg/mL CHX for 0–8 h as indicated. The statistical results of (<bold>C</bold>–<bold>E</bold>) are summarized in (<bold>E</bold>). Cell lysates were extracted and immunoblotted with respective antibodies. The intensity of the bands was quantified using LICOR Odyssey software and represented by trend lines. The experiments were repeated three times (n = 3) and representative images are presented. The representative western blot images are from different gels and each lane was loaded with the same amount of proteins. Data of triplicate assays are presented as mean ± S.D. * <italic>p</italic> <0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001; NS, no significance. “+” indicated that the plasmid was transfected, “−” indicated that the plasmid was not transfected.</p></caption><graphic xlink:href="biomolecules-09-00151-g004"></graphic></fig><fig id="biomolecules-09-00151-f005" orientation="portrait" position="float"><label>Figure 5</label><caption><p>SUMO pathway did not regulate DAPK1 protein levels via direct conjugation. (<bold>A</bold>) A diagram illustrating the panel of DAPK1 (1–364) point mutants. (<bold>B</bold>) HEK293T cells were transfected with LacZ, SENP6 and the DAPK1 point mutants as indicated. (<bold>C</bold>–<bold>E</bold>) HEK293T transfected with LacZ, DAPK1 (1–364) point mutants were exposed to 20 μg/mL CHX for 0–8 h as indicated and the statistical results were summarized in (<bold>F</bold>). The intensity of the bands was quantified using LICOR Odyssey software and represented by bar graphs. The experiments were repeated three times (n = 3) and representative images are presented. The representative western blot images are from different gels and each lane was loaded with the same amount of proteins. Data of triplicate assays are presented as mean ± S.D. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001; NS, no significance. “+” indicated that the plasmid was transfected, “−” indicated that the plasmid was not transfected.</p></caption><graphic xlink:href="biomolecules-09-00151-g005"></graphic></fig></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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