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PMID: 1063393 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Intracellular localization of mouse DNA polymerase-alpha.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 73 ·No. 4 ·1976-04-00 ·Pages 1136-9

Herrick G, Spear BB, Veomett G

Abstract

Although DNA polymerase-alpha (DNA nucleotidyltransferase; deoxynucleoside triphosphate: DNA deoxynucleotidyltransferase; EC 2.7.7.7) probably functions in the nucleus, it is usually found predominantly in the nonnuclear fraction of disrupted cells. We have reexamined the intracellular location of this enzyme using cytochalasin-B-induced enucleation, a technique which avoids exposure of nuclei to extra-cellular conditions during cell fractionation. In conditions where viability of separated cell parts is high and recovery is quantitative, we find greater than 85% of total DNA polymerase-alpha (and DNA polymerase-beta) activity in the nucleated cell fragments (karyoplasts), from which we conclude that the location in vivo of DNA polymerase-alpha is either nuclear or perinuclear. On the other hand, thymidine kinase (ATP: thymidine 5'-phosphotransferase, EC 5.7.1.75) is found primarily in the enucleated cell fragments (cytoplasts). The enucleation procedure used in this work should be of general use for intracellular location studies.

MeSH Terms
Cell Fractionation Cell Line Cell Nucleus/enzymology Chromatin/enzymology Cytochalasin B/pharmacology Cytoplasm/enzymology DNA Nucleotidyltransferases/metabolism Thymidine Kinase/metabolism
Chemicals
Chromatin Cytochalasin B Thymidine Kinase DNA Nucleotidyltransferases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Herrick G
Spear B B
Veomett G
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33 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.73.4.1136
Published
1976-04-00
Pages
1136-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC430215
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.73.4.1136 PMC Full Text View journal details
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