Cell-Cycle Dependence and Properties of the HeLa Cell DNA Polymerase System
Analysis of the properties of the DNA polymerase (pol) system as a function of fundamental factors of the assay environment allowed a rather accurate estimation of its dependence on the HeLa cell cycle. For pol α , the temperature and pH optima were 38.1 degrees C and 8.0, respectively; for pol β ,...
Gespeichert in:
| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1985-04, Vol.82 (8), p.2220-2224 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2224 |
|---|---|
| container_issue | 8 |
| container_start_page | 2220 |
| container_title | Proceedings of the National Academy of Sciences - PNAS |
| container_volume | 82 |
| creator | Delfini, Carlo Alfani, Elena De Venezia, Valeria Oberholtzer, Goffredo Tomasello, Carmelo Eremenko, Tamilla Volpe, Pietro |
| description | Analysis of the properties of the DNA polymerase (pol) system as a function of fundamental factors of the assay environment allowed a rather accurate estimation of its dependence on the HeLa cell cycle. For pol α , the temperature and pH optima were 38.1 degrees C and 8.0, respectively; for pol β , these optima were 36.2 degrees C and pH 7.4. Pol γ showed a pH optimum at 7.7. Optimum activity for both the α and β enzymes was observed at 60 mM Tris. The maximal activity at 36.2 degrees C and pH 7.4 was associated with resistance to N-ethylmaleimide (MalNEt), whereas that at 38.1 degrees C and pH 8.0 was sensitive to MalNEt. Incorporation of [3H]dTTP was maximal after 1 hr of incubation for the former activity and after 4 hr, for the latter. In extracts from cells in early S phase, the pol activity decreased after 1 hr of incubation, was MalNEt-resistant, and was characterized by temperature and pH optima at 36.2 degrees C and 7.4, respectively. In extracts of late S-phase cells, the polcatalyzed incorporation of [3H]dTTP continued after 4 hr of incubation, was MalNEt-sensitive, and was characterized by temperature and pH optima at 38.1 degrees C and 8.0, respectively. Thus, a pol β -type activity appeared in early S phase, whereas a pol α -type activity appeared in late S. During the G1, M, and G2phases, a background level of pol activity was observed that showed intermediate kinetic properties. |
| doi_str_mv | 10.1073/pnas.82.8.2220 |
| format | Article |
| fullrecord | <record><control><sourceid>jstor_pnas_</sourceid><recordid>TN_cdi_pnas_primary_82_8_2220_fulltext</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><jstor_id>25069</jstor_id><sourcerecordid>25069</sourcerecordid><originalsourceid>FETCH-LOGICAL-c516t-f6837ef2d1b2567a14c678ef9ff037e47a868824a6f150fe22ae5f93fe410b883</originalsourceid><addsrcrecordid>eNqFkc1rFDEYxoModVu9ehCEHKS3Gd9kMvk4eCjbaqWLFtRzyM6-sVPmq0lWuv-9M-w6rCCUHHL4Pc_79RDyhkHOQBUfhs7FXPNc55xzeEYWDAzLpDDwnCwAuMq04OIlOY3xHgBMqeGEnBS6VONbkJslNk223FUN0kscsNtgVyF13Ybehn7AkGqMtPc03SG9xpWjk4Fefr2gt32zazG4iPT7LiZsX5EX3jURXx_-M_Lz09WP5XW2-vb5y_JilVUlkynzUhcKPd-wNS-lckxUUmn0xnsYgVBOS625cNKzEjxy7rD0pvAoGKy1Ls7Ix33dYbtucVNhl4Jr7BDq1oWd7V1t_yVdfWd_9b9tYVTJJ__5wR_6hy3GZNs6VuNarsN-G62S42U1wJNCJgpRSGVGYb4XVqGPMaCfh2Fgp5jsFJPV3Go7xTQa3h2vMMsPuYz8_YG7WLnGB9dVdZxlhgnOhDoacCr_l85trN82TcLHdNTvv8KRv93z-5j6MAt4CdIUfwAo6rrd</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>14343679</pqid></control><display><type>article</type><title>Cell-Cycle Dependence and Properties of the HeLa Cell DNA Polymerase System</title><source>MEDLINE</source><source>Jstor Complete Legacy</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>Free Full-Text Journals in Chemistry</source><creator>Delfini, Carlo ; Alfani, Elena ; De Venezia, Valeria ; Oberholtzer, Goffredo ; Tomasello, Carmelo ; Eremenko, Tamilla ; Volpe, Pietro</creator><creatorcontrib>Delfini, Carlo ; Alfani, Elena ; De Venezia, Valeria ; Oberholtzer, Goffredo ; Tomasello, Carmelo ; Eremenko, Tamilla ; Volpe, Pietro</creatorcontrib><description>Analysis of the properties of the DNA polymerase (pol) system as a function of fundamental factors of the assay environment allowed a rather accurate estimation of its dependence on the HeLa cell cycle. For pol α , the temperature and pH optima were 38.1 degrees C and 8.0, respectively; for pol β , these optima were 36.2 degrees C and pH 7.4. Pol γ showed a pH optimum at 7.7. Optimum activity for both the α and β enzymes was observed at 60 mM Tris. The maximal activity at 36.2 degrees C and pH 7.4 was associated with resistance to N-ethylmaleimide (MalNEt), whereas that at 38.1 degrees C and pH 8.0 was sensitive to MalNEt. Incorporation of [3H]dTTP was maximal after 1 hr of incubation for the former activity and after 4 hr, for the latter. In extracts from cells in early S phase, the pol activity decreased after 1 hr of incubation, was MalNEt-resistant, and was characterized by temperature and pH optima at 36.2 degrees C and 7.4, respectively. In extracts of late S-phase cells, the polcatalyzed incorporation of [3H]dTTP continued after 4 hr of incubation, was MalNEt-sensitive, and was characterized by temperature and pH optima at 38.1 degrees C and 8.0, respectively. Thus, a pol β -type activity appeared in early S phase, whereas a pol α -type activity appeared in late S. During the G1, M, and G2phases, a background level of pol activity was observed that showed intermediate kinetic properties.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.82.8.2220</identifier><identifier>PMID: 3857575</identifier><identifier>CODEN: PNASA6</identifier><language>eng</language><publisher>Washington, DC: National Academy of Sciences of the United States of America</publisher><subject>Biochemistry ; Biological and medical sciences ; Cell Cycle ; Cell cycle, cell proliferation ; Cell physiology ; Cells ; DNA ; DNA Polymerase I - metabolism ; DNA Polymerase II - metabolism ; DNA Polymerase III - metabolism ; DNA-Directed DNA Polymerase - metabolism ; Enzymes ; Ethylmaleimide - pharmacology ; Fluorides ; Fundamental and applied biological sciences. Psychology ; HeLa cells ; HeLa Cells - cytology ; HeLa Cells - enzymology ; Humans ; Hydrogen-Ion Concentration ; Interphase ; Kinetics ; Legends ; Molecular and cellular biology ; Nucleic Acid Synthesis Inhibitors ; Temperature</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1985-04, Vol.82 (8), p.2220-2224</ispartof><rights>1985 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c516t-f6837ef2d1b2567a14c678ef9ff037e47a868824a6f150fe22ae5f93fe410b883</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/82/8.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/25069$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/25069$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,725,778,782,801,883,27907,27908,53774,53776,58000,58233</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9142147$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3857575$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Delfini, Carlo</creatorcontrib><creatorcontrib>Alfani, Elena</creatorcontrib><creatorcontrib>De Venezia, Valeria</creatorcontrib><creatorcontrib>Oberholtzer, Goffredo</creatorcontrib><creatorcontrib>Tomasello, Carmelo</creatorcontrib><creatorcontrib>Eremenko, Tamilla</creatorcontrib><creatorcontrib>Volpe, Pietro</creatorcontrib><title>Cell-Cycle Dependence and Properties of the HeLa Cell DNA Polymerase System</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Analysis of the properties of the DNA polymerase (pol) system as a function of fundamental factors of the assay environment allowed a rather accurate estimation of its dependence on the HeLa cell cycle. For pol α , the temperature and pH optima were 38.1 degrees C and 8.0, respectively; for pol β , these optima were 36.2 degrees C and pH 7.4. Pol γ showed a pH optimum at 7.7. Optimum activity for both the α and β enzymes was observed at 60 mM Tris. The maximal activity at 36.2 degrees C and pH 7.4 was associated with resistance to N-ethylmaleimide (MalNEt), whereas that at 38.1 degrees C and pH 8.0 was sensitive to MalNEt. Incorporation of [3H]dTTP was maximal after 1 hr of incubation for the former activity and after 4 hr, for the latter. In extracts from cells in early S phase, the pol activity decreased after 1 hr of incubation, was MalNEt-resistant, and was characterized by temperature and pH optima at 36.2 degrees C and 7.4, respectively. In extracts of late S-phase cells, the polcatalyzed incorporation of [3H]dTTP continued after 4 hr of incubation, was MalNEt-sensitive, and was characterized by temperature and pH optima at 38.1 degrees C and 8.0, respectively. Thus, a pol β -type activity appeared in early S phase, whereas a pol α -type activity appeared in late S. During the G1, M, and G2phases, a background level of pol activity was observed that showed intermediate kinetic properties.</description><subject>Biochemistry</subject><subject>Biological and medical sciences</subject><subject>Cell Cycle</subject><subject>Cell cycle, cell proliferation</subject><subject>Cell physiology</subject><subject>Cells</subject><subject>DNA</subject><subject>DNA Polymerase I - metabolism</subject><subject>DNA Polymerase II - metabolism</subject><subject>DNA Polymerase III - metabolism</subject><subject>DNA-Directed DNA Polymerase - metabolism</subject><subject>Enzymes</subject><subject>Ethylmaleimide - pharmacology</subject><subject>Fluorides</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HeLa cells</subject><subject>HeLa Cells - cytology</subject><subject>HeLa Cells - enzymology</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>Interphase</subject><subject>Kinetics</subject><subject>Legends</subject><subject>Molecular and cellular biology</subject><subject>Nucleic Acid Synthesis Inhibitors</subject><subject>Temperature</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1985</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1rFDEYxoModVu9ehCEHKS3Gd9kMvk4eCjbaqWLFtRzyM6-sVPmq0lWuv-9M-w6rCCUHHL4Pc_79RDyhkHOQBUfhs7FXPNc55xzeEYWDAzLpDDwnCwAuMq04OIlOY3xHgBMqeGEnBS6VONbkJslNk223FUN0kscsNtgVyF13Ybehn7AkGqMtPc03SG9xpWjk4Fefr2gt32zazG4iPT7LiZsX5EX3jURXx_-M_Lz09WP5XW2-vb5y_JilVUlkynzUhcKPd-wNS-lckxUUmn0xnsYgVBOS625cNKzEjxy7rD0pvAoGKy1Ls7Ix33dYbtucVNhl4Jr7BDq1oWd7V1t_yVdfWd_9b9tYVTJJ__5wR_6hy3GZNs6VuNarsN-G62S42U1wJNCJgpRSGVGYb4XVqGPMaCfh2Fgp5jsFJPV3Go7xTQa3h2vMMsPuYz8_YG7WLnGB9dVdZxlhgnOhDoacCr_l85trN82TcLHdNTvv8KRv93z-5j6MAt4CdIUfwAo6rrd</recordid><startdate>19850401</startdate><enddate>19850401</enddate><creator>Delfini, Carlo</creator><creator>Alfani, Elena</creator><creator>De Venezia, Valeria</creator><creator>Oberholtzer, Goffredo</creator><creator>Tomasello, Carmelo</creator><creator>Eremenko, Tamilla</creator><creator>Volpe, Pietro</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19850401</creationdate><title>Cell-Cycle Dependence and Properties of the HeLa Cell DNA Polymerase System</title><author>Delfini, Carlo ; Alfani, Elena ; De Venezia, Valeria ; Oberholtzer, Goffredo ; Tomasello, Carmelo ; Eremenko, Tamilla ; Volpe, Pietro</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c516t-f6837ef2d1b2567a14c678ef9ff037e47a868824a6f150fe22ae5f93fe410b883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1985</creationdate><topic>Biochemistry</topic><topic>Biological and medical sciences</topic><topic>Cell Cycle</topic><topic>Cell cycle, cell proliferation</topic><topic>Cell physiology</topic><topic>Cells</topic><topic>DNA</topic><topic>DNA Polymerase I - metabolism</topic><topic>DNA Polymerase II - metabolism</topic><topic>DNA Polymerase III - metabolism</topic><topic>DNA-Directed DNA Polymerase - metabolism</topic><topic>Enzymes</topic><topic>Ethylmaleimide - pharmacology</topic><topic>Fluorides</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HeLa cells</topic><topic>HeLa Cells - cytology</topic><topic>HeLa Cells - enzymology</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>Interphase</topic><topic>Kinetics</topic><topic>Legends</topic><topic>Molecular and cellular biology</topic><topic>Nucleic Acid Synthesis Inhibitors</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Delfini, Carlo</creatorcontrib><creatorcontrib>Alfani, Elena</creatorcontrib><creatorcontrib>De Venezia, Valeria</creatorcontrib><creatorcontrib>Oberholtzer, Goffredo</creatorcontrib><creatorcontrib>Tomasello, Carmelo</creatorcontrib><creatorcontrib>Eremenko, Tamilla</creatorcontrib><creatorcontrib>Volpe, Pietro</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Delfini, Carlo</au><au>Alfani, Elena</au><au>De Venezia, Valeria</au><au>Oberholtzer, Goffredo</au><au>Tomasello, Carmelo</au><au>Eremenko, Tamilla</au><au>Volpe, Pietro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cell-Cycle Dependence and Properties of the HeLa Cell DNA Polymerase System</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1985-04-01</date><risdate>1985</risdate><volume>82</volume><issue>8</issue><spage>2220</spage><epage>2224</epage><pages>2220-2224</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><coden>PNASA6</coden><abstract>Analysis of the properties of the DNA polymerase (pol) system as a function of fundamental factors of the assay environment allowed a rather accurate estimation of its dependence on the HeLa cell cycle. For pol α , the temperature and pH optima were 38.1 degrees C and 8.0, respectively; for pol β , these optima were 36.2 degrees C and pH 7.4. Pol γ showed a pH optimum at 7.7. Optimum activity for both the α and β enzymes was observed at 60 mM Tris. The maximal activity at 36.2 degrees C and pH 7.4 was associated with resistance to N-ethylmaleimide (MalNEt), whereas that at 38.1 degrees C and pH 8.0 was sensitive to MalNEt. Incorporation of [3H]dTTP was maximal after 1 hr of incubation for the former activity and after 4 hr, for the latter. In extracts from cells in early S phase, the pol activity decreased after 1 hr of incubation, was MalNEt-resistant, and was characterized by temperature and pH optima at 36.2 degrees C and 7.4, respectively. In extracts of late S-phase cells, the polcatalyzed incorporation of [3H]dTTP continued after 4 hr of incubation, was MalNEt-sensitive, and was characterized by temperature and pH optima at 38.1 degrees C and 8.0, respectively. Thus, a pol β -type activity appeared in early S phase, whereas a pol α -type activity appeared in late S. During the G1, M, and G2phases, a background level of pol activity was observed that showed intermediate kinetic properties.</abstract><cop>Washington, DC</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>3857575</pmid><doi>10.1073/pnas.82.8.2220</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0027-8424 |
| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1985-04, Vol.82 (8), p.2220-2224 |
| issn | 0027-8424 1091-6490 |
| language | eng |
| recordid | cdi_pnas_primary_82_8_2220_fulltext |
| source | MEDLINE; Jstor Complete Legacy; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Biochemistry Biological and medical sciences Cell Cycle Cell cycle, cell proliferation Cell physiology Cells DNA DNA Polymerase I - metabolism DNA Polymerase II - metabolism DNA Polymerase III - metabolism DNA-Directed DNA Polymerase - metabolism Enzymes Ethylmaleimide - pharmacology Fluorides Fundamental and applied biological sciences. Psychology HeLa cells HeLa Cells - cytology HeLa Cells - enzymology Humans Hydrogen-Ion Concentration Interphase Kinetics Legends Molecular and cellular biology Nucleic Acid Synthesis Inhibitors Temperature |
| title | Cell-Cycle Dependence and Properties of the HeLa Cell DNA Polymerase System |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-16T20%3A51%3A45IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-jstor_pnas_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Cell-Cycle%20Dependence%20and%20Properties%20of%20the%20HeLa%20Cell%20DNA%20Polymerase%20System&rft.jtitle=Proceedings%20of%20the%20National%20Academy%20of%20Sciences%20-%20PNAS&rft.au=Delfini,%20Carlo&rft.date=1985-04-01&rft.volume=82&rft.issue=8&rft.spage=2220&rft.epage=2224&rft.pages=2220-2224&rft.issn=0027-8424&rft.eissn=1091-6490&rft.coden=PNASA6&rft_id=info:doi/10.1073/pnas.82.8.2220&rft_dat=%3Cjstor_pnas_%3E25069%3C/jstor_pnas_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=14343679&rft_id=info:pmid/3857575&rft_jstor_id=25069&rfr_iscdi=true |