Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase.
Identifieur interne : 002502 ( PubMed/Corpus ); précédent : 002501; suivant : 002503Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase.
Auteurs : Kieu X. Luu ; Sreenivas Kanugula ; Anthony E. Pegg ; Gary T. Pauly ; Robert C. MoschelSource :
- Biochemistry [ 0006-2960 ] ; 2002.
English descriptors
- KwdEn :
- Alkaline Phosphatase (metabolism), Amino Acid Substitution, Animals, Base Sequence, Crotalus, DNA Repair, Escherichia coli (enzymology), Kinetics, Magnetic Resonance Spectroscopy, Mutagenesis, Site-Directed, O(6)-Methylguanine-DNA Methyltransferase (metabolism), Oligodeoxyribonucleotides (metabolism), Phosphodiesterase I, Phosphoric Diester Hydrolases, Recombinant Proteins (metabolism), Substrate Specificity.
- MESH :
- chemical , metabolism : Alkaline Phosphatase, O(6)-Methylguanine-DNA Methyltransferase, Oligodeoxyribonucleotides, Recombinant Proteins.
- enzymology : Escherichia coli.
- Amino Acid Substitution, Animals, Base Sequence, Crotalus, DNA Repair, Kinetics, Magnetic Resonance Spectroscopy, Mutagenesis, Site-Directed, Phosphodiesterase I, Phosphoric Diester Hydrolases, Substrate Specificity.
Abstract
Activity of the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT) is an important source of tumor cell resistance to alkylating agents. AGT inhibitors may prove useful in enhancing chemotherapy. AGT is inactivated by reacting stoichiometrically with O(6)-benzylguanine (b(6)G), which is currently in clinical trials for this purpose. Short oligodeoxyribonucleotides containing a central b(6)G are more potent inactivators of AGT than b(6)G. We examined whether human AGT could react with oligodeoxyribonucleotides containing multiple b(6)G residues. The single-stranded 7-mer 5'-d[T(b(6)G)(5)G]-3' was an excellent AGT substrate with all five b(6)G adducts repaired although one adduct was repaired much more slowly. The highly b(6)G-resistant Y158H and P140K AGT mutants were also inactivated by 5'-d[T(b(6)G)(5)G]-3'. Studies with 7-mers containing a single b(6)G adduct showed that 5'-d[TGGGG(b(6)G)G]-3' was more poorly repaired by wild-type AGT than 5'-d[T(b(6)G)GGGGG]-3' and 5'-d[TGG(b(6)G)GGG]-3' and was even less repairable by mutants Y158H and P140K. This positional effect was unaffected by interchanging the terminal 5'- or 3'-nucleotides and was also observed with single-stranded 16-mer oligodeoxyribonucleotides containing O(6)-methylguanine, where a minimum of four nucleotides 3' to the lesion was required for the most efficient repair. Annealing with the reverse complementary strands to produce double-stranded substrates increased the ability of AGT to repair adducts at all positions except at positions 2 and 15. Our results suggest that AGT recognizes the polarity of single-stranded DNA, with the best substrates having an adduct adjacent to the 5'-terminal residue. These findings will aid in designing novel AGT inhibitors that incorporate O(6)-alkylguanine adducts in oligodeoxyribonucleotide contexts.
DOI: 10.1021/bi025857i
PubMed: 12093287
Links to Exploration step
pubmed:12093287Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase.</title><author><name sortKey="Luu, Kieu X" sort="Luu, Kieu X" uniqKey="Luu K" first="Kieu X" last="Luu">Kieu X. Luu</name><affiliation><nlm:affiliation>Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, The Milton S. Hershey Medical Center, P.O. Box 850, 500 University Drive, Hershey, Pennsylvania 17033-0850, USA.</nlm:affiliation></affiliation></author><author><name sortKey="Kanugula, Sreenivas" sort="Kanugula, Sreenivas" uniqKey="Kanugula S" first="Sreenivas" last="Kanugula">Sreenivas Kanugula</name></author><author><name sortKey="Pegg, Anthony E" sort="Pegg, Anthony E" uniqKey="Pegg A" first="Anthony E" last="Pegg">Anthony E. Pegg</name></author><author><name sortKey="Pauly, Gary T" sort="Pauly, Gary T" uniqKey="Pauly G" first="Gary T" last="Pauly">Gary T. Pauly</name></author><author><name sortKey="Moschel, Robert C" sort="Moschel, Robert C" uniqKey="Moschel R" first="Robert C" last="Moschel">Robert C. Moschel</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2002">2002</date><idno type="RBID">pubmed:12093287</idno><idno type="pmid">12093287</idno><idno type="doi">10.1021/bi025857i</idno><idno type="wicri:Area/PubMed/Corpus">002502</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002502</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase.</title><author><name sortKey="Luu, Kieu X" sort="Luu, Kieu X" uniqKey="Luu K" first="Kieu X" last="Luu">Kieu X. Luu</name><affiliation><nlm:affiliation>Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, The Milton S. Hershey Medical Center, P.O. Box 850, 500 University Drive, Hershey, Pennsylvania 17033-0850, USA.</nlm:affiliation></affiliation></author><author><name sortKey="Kanugula, Sreenivas" sort="Kanugula, Sreenivas" uniqKey="Kanugula S" first="Sreenivas" last="Kanugula">Sreenivas Kanugula</name></author><author><name sortKey="Pegg, Anthony E" sort="Pegg, Anthony E" uniqKey="Pegg A" first="Anthony E" last="Pegg">Anthony E. Pegg</name></author><author><name sortKey="Pauly, Gary T" sort="Pauly, Gary T" uniqKey="Pauly G" first="Gary T" last="Pauly">Gary T. Pauly</name></author><author><name sortKey="Moschel, Robert C" sort="Moschel, Robert C" uniqKey="Moschel R" first="Robert C" last="Moschel">Robert C. Moschel</name></author></analytic><series><title level="j">Biochemistry</title><idno type="ISSN">0006-2960</idno><imprint><date when="2002" type="published">2002</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Alkaline Phosphatase (metabolism)</term><term>Amino Acid Substitution</term><term>Animals</term><term>Base Sequence</term><term>Crotalus</term><term>DNA Repair</term><term>Escherichia coli (enzymology)</term><term>Kinetics</term><term>Magnetic Resonance Spectroscopy</term><term>Mutagenesis, Site-Directed</term><term>O(6)-Methylguanine-DNA Methyltransferase (metabolism)</term><term>Oligodeoxyribonucleotides (metabolism)</term><term>Phosphodiesterase I</term><term>Phosphoric Diester Hydrolases</term><term>Recombinant Proteins (metabolism)</term><term>Substrate Specificity</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Alkaline Phosphatase</term><term>O(6)-Methylguanine-DNA Methyltransferase</term><term>Oligodeoxyribonucleotides</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Escherichia coli</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Substitution</term><term>Animals</term><term>Base Sequence</term><term>Crotalus</term><term>DNA Repair</term><term>Kinetics</term><term>Magnetic Resonance Spectroscopy</term><term>Mutagenesis, Site-Directed</term><term>Phosphodiesterase I</term><term>Phosphoric Diester Hydrolases</term><term>Substrate Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Activity of the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT) is an important source of tumor cell resistance to alkylating agents. AGT inhibitors may prove useful in enhancing chemotherapy. AGT is inactivated by reacting stoichiometrically with O(6)-benzylguanine (b(6)G), which is currently in clinical trials for this purpose. Short oligodeoxyribonucleotides containing a central b(6)G are more potent inactivators of AGT than b(6)G. We examined whether human AGT could react with oligodeoxyribonucleotides containing multiple b(6)G residues. The single-stranded 7-mer 5'-d[T(b(6)G)(5)G]-3' was an excellent AGT substrate with all five b(6)G adducts repaired although one adduct was repaired much more slowly. The highly b(6)G-resistant Y158H and P140K AGT mutants were also inactivated by 5'-d[T(b(6)G)(5)G]-3'. Studies with 7-mers containing a single b(6)G adduct showed that 5'-d[TGGGG(b(6)G)G]-3' was more poorly repaired by wild-type AGT than 5'-d[T(b(6)G)GGGGG]-3' and 5'-d[TGG(b(6)G)GGG]-3' and was even less repairable by mutants Y158H and P140K. This positional effect was unaffected by interchanging the terminal 5'- or 3'-nucleotides and was also observed with single-stranded 16-mer oligodeoxyribonucleotides containing O(6)-methylguanine, where a minimum of four nucleotides 3' to the lesion was required for the most efficient repair. Annealing with the reverse complementary strands to produce double-stranded substrates increased the ability of AGT to repair adducts at all positions except at positions 2 and 15. Our results suggest that AGT recognizes the polarity of single-stranded DNA, with the best substrates having an adduct adjacent to the 5'-terminal residue. These findings will aid in designing novel AGT inhibitors that incorporate O(6)-alkylguanine adducts in oligodeoxyribonucleotide contexts.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">12093287</PMID><DateCompleted><Year>2002</Year><Month>08</Month><Day>21</Day></DateCompleted><DateRevised><Year>2019</Year><Month>06</Month><Day>13</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0006-2960</ISSN><JournalIssue CitedMedium="Print"><Volume>41</Volume><Issue>27</Issue><PubDate><Year>2002</Year><Month>Jul</Month><Day>09</Day></PubDate></JournalIssue><Title>Biochemistry</Title><ISOAbbreviation>Biochemistry</ISOAbbreviation></Journal><ArticleTitle>Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase.</ArticleTitle><Pagination><MedlinePgn>8689-97</MedlinePgn></Pagination><Abstract><AbstractText>Activity of the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT) is an important source of tumor cell resistance to alkylating agents. AGT inhibitors may prove useful in enhancing chemotherapy. AGT is inactivated by reacting stoichiometrically with O(6)-benzylguanine (b(6)G), which is currently in clinical trials for this purpose. Short oligodeoxyribonucleotides containing a central b(6)G are more potent inactivators of AGT than b(6)G. We examined whether human AGT could react with oligodeoxyribonucleotides containing multiple b(6)G residues. The single-stranded 7-mer 5'-d[T(b(6)G)(5)G]-3' was an excellent AGT substrate with all five b(6)G adducts repaired although one adduct was repaired much more slowly. The highly b(6)G-resistant Y158H and P140K AGT mutants were also inactivated by 5'-d[T(b(6)G)(5)G]-3'. Studies with 7-mers containing a single b(6)G adduct showed that 5'-d[TGGGG(b(6)G)G]-3' was more poorly repaired by wild-type AGT than 5'-d[T(b(6)G)GGGGG]-3' and 5'-d[TGG(b(6)G)GGG]-3' and was even less repairable by mutants Y158H and P140K. This positional effect was unaffected by interchanging the terminal 5'- or 3'-nucleotides and was also observed with single-stranded 16-mer oligodeoxyribonucleotides containing O(6)-methylguanine, where a minimum of four nucleotides 3' to the lesion was required for the most efficient repair. Annealing with the reverse complementary strands to produce double-stranded substrates increased the ability of AGT to repair adducts at all positions except at positions 2 and 15. Our results suggest that AGT recognizes the polarity of single-stranded DNA, with the best substrates having an adduct adjacent to the 5'-terminal residue. These findings will aid in designing novel AGT inhibitors that incorporate O(6)-alkylguanine adducts in oligodeoxyribonucleotide contexts.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Luu</LastName><ForeName>Kieu X</ForeName><Initials>KX</Initials><AffiliationInfo><Affiliation>Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, The Milton S. Hershey Medical Center, P.O. Box 850, 500 University Drive, Hershey, Pennsylvania 17033-0850, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kanugula</LastName><ForeName>Sreenivas</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Pegg</LastName><ForeName>Anthony E</ForeName><Initials>AE</Initials></Author><Author ValidYN="Y"><LastName>Pauly</LastName><ForeName>Gary T</ForeName><Initials>GT</Initials></Author><Author ValidYN="Y"><LastName>Moschel</LastName><ForeName>Robert C</ForeName><Initials>RC</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>CA-18137</GrantID><Acronym>CA</Acronym><Agency>NCI NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>CA-71976</GrantID><Acronym>CA</Acronym><Agency>NCI NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biochemistry</MedlineTA><NlmUniqueID>0370623</NlmUniqueID><ISSNLinking>0006-2960</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009838">Oligodeoxyribonucleotides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.1.1.63</RegistryNumber><NameOfSubstance UI="D019853">O(6)-Methylguanine-DNA Methyltransferase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.1.3.1</RegistryNumber><NameOfSubstance UI="D000469">Alkaline Phosphatase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.1.4.-</RegistryNumber><NameOfSubstance UI="D010727">Phosphoric Diester Hydrolases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.1.4.1</RegistryNumber><NameOfSubstance UI="D043264">Phosphodiesterase I</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000469" MajorTopicYN="N">Alkaline Phosphatase</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019943" MajorTopicYN="N">Amino Acid Substitution</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017839" MajorTopicYN="N">Crotalus</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004260" MajorTopicYN="Y">DNA Repair</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009682" MajorTopicYN="N">Magnetic Resonance Spectroscopy</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016297" MajorTopicYN="N">Mutagenesis, Site-Directed</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019853" MajorTopicYN="N">O(6)-Methylguanine-DNA Methyltransferase</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009838" MajorTopicYN="N">Oligodeoxyribonucleotides</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D043264" MajorTopicYN="N">Phosphodiesterase I</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010727" MajorTopicYN="N">Phosphoric Diester Hydrolases</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013379" MajorTopicYN="N">Substrate Specificity</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2002</Year><Month>7</Month><Day>3</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2002</Year><Month>8</Month><Day>22</Day><Hour>10</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2002</Year><Month>7</Month><Day>3</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">12093287</ArticleId><ArticleId IdType="pii">bi025857i</ArticleId><ArticleId IdType="doi">10.1021/bi025857i</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/PubMed/Corpus
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002502 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/PubMed/Corpus/biblio.hfd -nk 002502 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= PubMed
|étape= Corpus
|type= RBID
|clé= pubmed:12093287
|texte= Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/PubMed/Corpus/RBID.i -Sk "pubmed:12093287" \
| HfdSelect -Kh $EXPLOR_AREA/Data/PubMed/Corpus/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |