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PMID: 3025653 Published · ppublish English Journal Article

Immortalization of rat embryo fibroblasts by mutant polyomavirus large T antigens deficient in DNA binding.

Molecular and cellular biology ·Vol. 6 ·No. 12 ·1986-12-00 ·Pages 4344-52

Cowie A, de Villiers J, Kamen R

Abstract

We have identified a putative DNA-binding domain in polyomavirus large T antigen. Mutations introduced into the gene between amino acids 290 and 310 resulted in proteins that no longer bound to the high-affinity binding sites on the polyomavirus genome, showed no detectable nonspecific DNA binding, and were not able to initiate DNA replication from the viral origin. These mutant T antigen genes were introduced into rat embryo fibroblasts together with the neomycin resistance gene to allow selection for growth in the presence of G418. All the mutations tested facilitated the establishment of these cells in long-term culture at an efficiency indistinguishable from that of the wild-type protein.

MeSH Terms
Amino Acid Sequence Animals Antigens, Polyomavirus Transforming Antigens, Viral, Tumor/genetics,metabolism Cats Cell Line DNA, Viral/metabolism DNA-Binding Proteins Embryo, Mammalian Genes Genes, Viral Mutation Oncogene Proteins, Viral/genetics,metabolism Plasmids Polyomavirus/genetics Protein Kinases/genetics
Chemicals
Antigens, Polyomavirus Transforming Antigens, Viral, Tumor DNA, Viral DNA-Binding Proteins Oncogene Proteins, Viral Protein Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Cowie A
de Villiers J
Kamen R
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55 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.12.4344-4352.1986
Published
1986-12-00
Pages
4344-52
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367216
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.6.12.4344-4352.1986 PMC Full Text View journal details
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