Mechanism of cellular uptake of modified oligodeoxynucleotides containing methylphosphonate linkages.
Identifieur interne : 002977 ( PubMed/Corpus ); précédent : 002976; suivant : 002978Mechanism of cellular uptake of modified oligodeoxynucleotides containing methylphosphonate linkages.
Auteurs : Y. Shoji ; S. Akhtar ; A. Periasamy ; B. Herman ; R L JulianoSource :
- Nucleic acids research [ 0305-1048 ] ; 1991.
English descriptors
- KwdEn :
- Animals, Base Sequence, Binding, Competitive, Biological Transport, Cell Line, Cell Membrane (metabolism), Cricetinae, Electrophoresis, Polyacrylamide Gel, Endocytosis, Humans, Hydrogen-Ion Concentration, Image Processing, Computer-Assisted, Kinetics, Molecular Sequence Data, Oligodeoxyribonucleotides (chemistry), Oligodeoxyribonucleotides (metabolism), Organophosphorus Compounds (chemistry), Organophosphorus Compounds (metabolism), Phosphorus Radioisotopes, Receptors, Cell Surface (antagonists & inhibitors), Rhodamines, Temperature.
- MESH :
- chemical , antagonists & inhibitors : Receptors, Cell Surface.
- chemical , chemistry : Oligodeoxyribonucleotides, Organophosphorus Compounds.
- metabolism : Cell Membrane, Oligodeoxyribonucleotides, Organophosphorus Compounds.
- Animals, Base Sequence, Binding, Competitive, Biological Transport, Cell Line, Cricetinae, Electrophoresis, Polyacrylamide Gel, Endocytosis, Humans, Hydrogen-Ion Concentration, Image Processing, Computer-Assisted, Kinetics, Molecular Sequence Data, Phosphorus Radioisotopes, Rhodamines, Temperature.
Abstract
The cellular uptake and intracellular distribution of methylphosphonate oligonucleotides (15 mers) has been examined using both 32P labeled and fluorescent labeled oligonucleotides. The cellular uptake process for methylphosphonate oligonucleotides is highly temperature dependent, with a major increase in uptake occurring between 15 and 20 degrees C. Most of the label which becomes cell associated at 37 degrees C cannot be removed by acid washing or trypsinization and thus seems to be within the cell. Visualization of rhodamine labeled methylphosphonate oligonucleotides using digital imaging fluorescence microscopy reveals a vesicular subcellular distribution suggestive of an endosomal localization. There was extensive co-localization of rhodamine labeled methylphosphonate oligonucleotides with fluorescein-dextran, an endosomal/lysosomal marker substance. The apparent endocytotic uptake of labeled methylphosphonate oligonucleotides could not be blocked by competition with unlabeled methylphosphonate or phosphodiester oligonucleotides, nor by ATP. This contrasts with the situation for radiolabeled phosphodiester oligonucleotides whose uptake can be completely blocked with unlabeled competitor. Uptake of phosphodiester oligonucleotides, but not of methylphosphonate oligonucleotides, could be blocked by acidification of the cytosol. These observations suggest that the pathway of cellular uptake of methylphosphonate oligonucleotides involves fluid phase or adsorbtive endocytosis, and is distinct from the uptake pathway for phosphodiester oligonucleotides.
DOI: 10.1093/nar/19.20.5543
PubMed: 1658734
Links to Exploration step
pubmed:1658734Le document en format XML
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Herman</name></author><author><name sortKey="Juliano, R L" sort="Juliano, R L" uniqKey="Juliano R" first="R L" last="Juliano">R L Juliano</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1991">1991</date><idno type="RBID">pubmed:1658734</idno><idno type="pmid">1658734</idno><idno type="doi">10.1093/nar/19.20.5543</idno><idno type="wicri:Area/PubMed/Corpus">002977</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002977</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Mechanism of cellular uptake of modified oligodeoxynucleotides containing methylphosphonate linkages.</title><author><name sortKey="Shoji, Y" sort="Shoji, Y" uniqKey="Shoji Y" first="Y" last="Shoji">Y. Shoji</name><affiliation><nlm:affiliation>Department of Pharmacology, University of North Carolina, Chapel Hill 27599.</nlm:affiliation></affiliation></author><author><name sortKey="Akhtar, S" sort="Akhtar, S" uniqKey="Akhtar S" first="S" last="Akhtar">S. Akhtar</name></author><author><name sortKey="Periasamy, A" sort="Periasamy, A" uniqKey="Periasamy A" first="A" last="Periasamy">A. Periasamy</name></author><author><name sortKey="Herman, B" sort="Herman, B" uniqKey="Herman B" first="B" last="Herman">B. Herman</name></author><author><name sortKey="Juliano, R L" sort="Juliano, R L" uniqKey="Juliano R" first="R L" last="Juliano">R L Juliano</name></author></analytic><series><title level="j">Nucleic acids research</title><idno type="ISSN">0305-1048</idno><imprint><date when="1991" type="published">1991</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Binding, Competitive</term><term>Biological Transport</term><term>Cell Line</term><term>Cell Membrane (metabolism)</term><term>Cricetinae</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Endocytosis</term><term>Humans</term><term>Hydrogen-Ion Concentration</term><term>Image Processing, Computer-Assisted</term><term>Kinetics</term><term>Molecular Sequence Data</term><term>Oligodeoxyribonucleotides (chemistry)</term><term>Oligodeoxyribonucleotides (metabolism)</term><term>Organophosphorus Compounds (chemistry)</term><term>Organophosphorus Compounds (metabolism)</term><term>Phosphorus Radioisotopes</term><term>Receptors, Cell Surface (antagonists & inhibitors)</term><term>Rhodamines</term><term>Temperature</term></keywords><keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>Receptors, Cell Surface</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Oligodeoxyribonucleotides</term><term>Organophosphorus Compounds</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Cell Membrane</term><term>Oligodeoxyribonucleotides</term><term>Organophosphorus Compounds</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Binding, Competitive</term><term>Biological Transport</term><term>Cell Line</term><term>Cricetinae</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Endocytosis</term><term>Humans</term><term>Hydrogen-Ion Concentration</term><term>Image Processing, Computer-Assisted</term><term>Kinetics</term><term>Molecular Sequence Data</term><term>Phosphorus Radioisotopes</term><term>Rhodamines</term><term>Temperature</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The cellular uptake and intracellular distribution of methylphosphonate oligonucleotides (15 mers) has been examined using both 32P labeled and fluorescent labeled oligonucleotides. The cellular uptake process for methylphosphonate oligonucleotides is highly temperature dependent, with a major increase in uptake occurring between 15 and 20 degrees C. Most of the label which becomes cell associated at 37 degrees C cannot be removed by acid washing or trypsinization and thus seems to be within the cell. Visualization of rhodamine labeled methylphosphonate oligonucleotides using digital imaging fluorescence microscopy reveals a vesicular subcellular distribution suggestive of an endosomal localization. There was extensive co-localization of rhodamine labeled methylphosphonate oligonucleotides with fluorescein-dextran, an endosomal/lysosomal marker substance. The apparent endocytotic uptake of labeled methylphosphonate oligonucleotides could not be blocked by competition with unlabeled methylphosphonate or phosphodiester oligonucleotides, nor by ATP. This contrasts with the situation for radiolabeled phosphodiester oligonucleotides whose uptake can be completely blocked with unlabeled competitor. Uptake of phosphodiester oligonucleotides, but not of methylphosphonate oligonucleotides, could be blocked by acidification of the cytosol. These observations suggest that the pathway of cellular uptake of methylphosphonate oligonucleotides involves fluid phase or adsorbtive endocytosis, and is distinct from the uptake pathway for phosphodiester oligonucleotides.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">1658734</PMID><DateCompleted><Year>1991</Year><Month>12</Month><Day>02</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>01</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0305-1048</ISSN><JournalIssue CitedMedium="Print"><Volume>19</Volume><Issue>20</Issue><PubDate><Year>1991</Year><Month>Oct</Month><Day>25</Day></PubDate></JournalIssue><Title>Nucleic acids research</Title><ISOAbbreviation>Nucleic Acids Res.</ISOAbbreviation></Journal><ArticleTitle>Mechanism of cellular uptake of modified oligodeoxynucleotides containing methylphosphonate linkages.</ArticleTitle><Pagination><MedlinePgn>5543-50</MedlinePgn></Pagination><Abstract><AbstractText>The cellular uptake and intracellular distribution of methylphosphonate oligonucleotides (15 mers) has been examined using both 32P labeled and fluorescent labeled oligonucleotides. The cellular uptake process for methylphosphonate oligonucleotides is highly temperature dependent, with a major increase in uptake occurring between 15 and 20 degrees C. Most of the label which becomes cell associated at 37 degrees C cannot be removed by acid washing or trypsinization and thus seems to be within the cell. Visualization of rhodamine labeled methylphosphonate oligonucleotides using digital imaging fluorescence microscopy reveals a vesicular subcellular distribution suggestive of an endosomal localization. There was extensive co-localization of rhodamine labeled methylphosphonate oligonucleotides with fluorescein-dextran, an endosomal/lysosomal marker substance. The apparent endocytotic uptake of labeled methylphosphonate oligonucleotides could not be blocked by competition with unlabeled methylphosphonate or phosphodiester oligonucleotides, nor by ATP. This contrasts with the situation for radiolabeled phosphodiester oligonucleotides whose uptake can be completely blocked with unlabeled competitor. Uptake of phosphodiester oligonucleotides, but not of methylphosphonate oligonucleotides, could be blocked by acidification of the cytosol. These observations suggest that the pathway of cellular uptake of methylphosphonate oligonucleotides involves fluid phase or adsorbtive endocytosis, and is distinct from the uptake pathway for phosphodiester oligonucleotides.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Shoji</LastName><ForeName>Y</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Department of Pharmacology, University of North Carolina, Chapel Hill 27599.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Akhtar</LastName><ForeName>S</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Periasamy</LastName><ForeName>A</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Herman</LastName><ForeName>B</ForeName><Initials>B</Initials></Author><Author ValidYN="Y"><LastName>Juliano</LastName><ForeName>R L</ForeName><Initials>RL</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>CA47044</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>England</Country><MedlineTA>Nucleic Acids Res</MedlineTA><NlmUniqueID>0411011</NlmUniqueID><ISSNLinking>0305-1048</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009838">Oligodeoxyribonucleotides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009943">Organophosphorus Compounds</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010761">Phosphorus Radioisotopes</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance 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