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. 1991 Dec;65(12):6998–7003. doi: 10.1128/jvi.65.12.6998-7003.1991

Determination of the origin-specific DNA-binding domain of polyomavirus large T antigen.

N A Sunstrom 1, N H Acheson 1, J A Hassell 1
PMCID: PMC250815  PMID: 1658390

Abstract

To map the DNA-binding domain of polyomavirus large T antigen, we constructed a set of plasmids coding for unidirectional carboxy- or amino-terminal deletion mutations in the large T antigen. Analysis of origin-specific DNA binding by mutant proteins expressed in Cos-1 cells revealed that the C-terminal boundary of the DNA-binding domain is at or near Glu-398. Fusion proteins of large T antigen lacking the first 200 N-terminal amino acids bound specifically to polyomavirus origin DNA; however, deletions beyond this site resulted in unstable proteins which could not be tested for DNA binding. Testing of point mutants and internal deletions by others suggested that the N-terminal boundary of the DNA-binding domain lies between amino acids 282 and 286. Taken together, these results locate the DNA-binding domain of polyomavirus large T antigen to the 116-amino-acid region between residues 282 and 398.

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Selected References

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