The sequences between 0.59 and 0.54 map units on SV40 DNA code for the unique region of small t antigen

Paucha, E.; Smith, A.E.

Cell 15(3): 1011-1020

1978


ISSN/ISBN: 0092-8674
PMID: 215314
DOI: 10.1016/0092-8674(78)90285-4
Accession: 068524358

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Abstract
To demonstrate directly that the carboxy terminal portion of SV-40 small t cells, or from cell-free systems primed with mRNA from mutant-infected cells, resulted in only low yields of the fragments. Experiments using purified SV-40 mRNA in low background cell-free systems, in which large T and small t could be detected without immunoprecipitation, suggested that these low yields were accounted for by reduced amounts of mRNA coding for the shortened forms of small t present in the mutant-infected cells. Larger amounts of putative fragments of small t were produced by translation of deletion mutant cRNA (complementary RNA synthesized in vitro using purified deletion mutant DNA and Escherichia coli RNA polymerase). Fingerprint analysis of the proteins produced showed that they contain most, if not all, of the methionine peptides common to small t and large T. The fragments of small t produced in response to dl 884 and dl 890 lack 2 methionine peptides that are present in small t but not in large T. These data provide direct evidence that the region between 0.59-0.54 map units on SV-40 DNA codes for polypeptide sequences that are unique to small t, and establishes that the nucleotide sequences from the region between 0.59-0.54 map units are both a coding sequence (for small t) and an intervening sequence (for large T).