Purification of an adenovirus-coded DNA polymerase that is required for initiation of DNA replication

Stillman, B. W., Tamanoi, F., Mathews, M. B. (1982) Purification of an adenovirus-coded DNA polymerase that is required for initiation of DNA replication. Cell, 31 (3 Pt 2). pp. 613-23. ISSN 0092-8674 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/6297776
DOI: 10.1016/0092-8674(82)90317-8

Abstract

Temperature-sensitive mutants in the N complementation group of human adenovirus type 5 are defective at the nonpermissive temperature for replication of virus DNA and for transformation of rat embryo cells. We show that nuclear extracts prepared from Ad5ts 149-infected cells grown at the nonpermissive temperature fail to replicate DNA in vitro. The defect lies in the first step in the initiation of viral DNA synthesis, the formation of a covalent linkage between the terminal protein precursor (pTP) and dCMP. A 140 kilodalton (140 kd) protein which complements these defective extracts and contains DNA polymerase activity has been purified from HeLa cells infected with wild-type Ad2. It is tightly associated with the 80 kd pTP in a replication complex. Both of these proteins are products of the E2B region of the adenovirus genome, and the 140 kd protein coding sequences lie immediately downstream from those encoding the 80 kd protein. These results demonstrate that adenovirus encodes a novel DNA polymerase that is required for priming of DNA synthesis at the origin of replication. This protein may also function in the initiation of transformation of cultured cells.

Item Type: Paper
Uncontrolled Keywords: Adenoviruses, Human enzymology genetics Cell Transformation, Neoplastic DNA Replication DNA Restriction Enzymes DNA-Directed DNA Polymerase genetics isolation & purification Genes Genes, Viral Hela Cells enzymology Humans Protein Biosynthesis RNA, Messenger genetics Transcription Genetic
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
organism description > virus
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 29 Feb 2012 14:18
Last Modified: 20 Jun 2017 20:43
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25108

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