Mechanism of cellular uptake of modified oligodeoxynucleotides containing methylphosphonate linkages.
Identifieur interne : 004870 ( Main/Exploration ); précédent : 004869; suivant : 004871Mechanism 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.
Descripteurs français
- KwdFr :
- Animaux, Cinétique, Composés organiques du phosphore (), Composés organiques du phosphore (métabolisme), Concentration en ions d'hydrogène, Cricetinae, Données de séquences moléculaires, Endocytose, Fixation compétitive, Humains, Lignée cellulaire, Membrane cellulaire (métabolisme), Oligodésoxyribonucléotides (), Oligodésoxyribonucléotides (métabolisme), Radio-isotopes du phosphore, Rhodamines, Récepteurs de surface cellulaire (antagonistes et inhibiteurs), Séquence nucléotidique, Température, Traitement d'image par ordinateur, Transport biologique, Électrophorèse sur gel de polyacrylamide.
- MESH :
- antagonistes et inhibiteurs : Récepteurs de surface cellulaire.
- métabolisme : Composés organiques du phosphore, Membrane cellulaire, Oligodésoxyribonucléotides.
- Animaux, Cinétique, Composés organiques du phosphore, Concentration en ions d'hydrogène, Cricetinae, Données de séquences moléculaires, Endocytose, Fixation compétitive, Humains, Lignée cellulaire, Oligodésoxyribonucléotides, Radio-isotopes du phosphore, Rhodamines, Séquence nucléotidique, Température, Traitement d'image par ordinateur, Transport biologique, Électrophorèse sur gel de polyacrylamide.
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
Affiliations:
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Shoji</name><affiliation><nlm:affiliation>Department of Pharmacology, University of North Carolina, Chapel Hill 27599.</nlm:affiliation><wicri:noCountry code="subField">Chapel Hill 27599</wicri:noCountry></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="KwdFr" xml:lang="fr"><term>Animaux</term><term>Cinétique</term><term>Composés organiques du phosphore ()</term><term>Composés organiques du phosphore (métabolisme)</term><term>Concentration en ions d'hydrogène</term><term>Cricetinae</term><term>Données de séquences moléculaires</term><term>Endocytose</term><term>Fixation compétitive</term><term>Humains</term><term>Lignée cellulaire</term><term>Membrane cellulaire (métabolisme)</term><term>Oligodésoxyribonucléotides ()</term><term>Oligodésoxyribonucléotides (métabolisme)</term><term>Radio-isotopes du phosphore</term><term>Rhodamines</term><term>Récepteurs de surface cellulaire (antagonistes et inhibiteurs)</term><term>Séquence nucléotidique</term><term>Température</term><term>Traitement d'image par ordinateur</term><term>Transport biologique</term><term>Électrophorèse sur gel de polyacrylamide</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="antagonistes et inhibiteurs" xml:lang="fr"><term>Récepteurs de surface cellulaire</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" qualifier="métabolisme" xml:lang="fr"><term>Composés organiques du phosphore</term><term>Membrane cellulaire</term><term>Oligodésoxyribonucléotides</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><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cinétique</term><term>Composés organiques du phosphore</term><term>Concentration en ions d'hydrogène</term><term>Cricetinae</term><term>Données de séquences moléculaires</term><term>Endocytose</term><term>Fixation compétitive</term><term>Humains</term><term>Lignée cellulaire</term><term>Oligodésoxyribonucléotides</term><term>Radio-isotopes du phosphore</term><term>Rhodamines</term><term>Séquence nucléotidique</term><term>Température</term><term>Traitement d'image par ordinateur</term><term>Transport biologique</term><term>Électrophorèse sur gel de polyacrylamide</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><affiliations><list></list><tree><noCountry><name sortKey="Akhtar, S" sort="Akhtar, S" uniqKey="Akhtar S" first="S" last="Akhtar">S. Akhtar</name><name sortKey="Herman, B" sort="Herman, B" uniqKey="Herman B" first="B" last="Herman">B. Herman</name><name sortKey="Juliano, R L" sort="Juliano, R L" uniqKey="Juliano R" first="R L" last="Juliano">R L Juliano</name><name sortKey="Periasamy, A" sort="Periasamy, A" uniqKey="Periasamy A" first="A" last="Periasamy">A. Periasamy</name><name sortKey="Shoji, Y" sort="Shoji, Y" uniqKey="Shoji Y" first="Y" last="Shoji">Y. Shoji</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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