Polyomavirus middle T-antigen NPTY mutants

Brian Druker, Lisa Sibert, Tmomas M. Roberts

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A polyomavirus middle T-antigen (MTAg) mutant containing a substitution of Leu for Pro at amino acid 248 has previously been described as completely transformation defective (B. J. Druker, L. Ling, B. Cohen, T. M. Roberts, and B. S. Schaffhausen, J. Virol. 64:4454-4461, 1990). This mutant had no alterations in associated proteins or associated kinase activities compared with wild-type MTAg. Pro-248 lies in a tetrametric sequence, NPTY, which is reminiscent of the so-called NPXY sequence in the low-density-lipoprotein receptor. In the low-density-lipoprotein receptor, mutations in the NPXY motif but not in the surrounding amino acids abolish receptor function, apparently by decreasing receptor internalization (W. Chen, J. L. Goldstein, and M. S. Brown, J. Biol. Chem. 265:3116-3123, 1990). To determine whether this sequence represents a functional motif in MTAg as well, a series of single amino acid substitutions was constructed in this region of MTAg. All of the mutations of N, P, T, or Y, including the relatively conservative substitution of Ser for Thr at amino acid 249, resulted in a transformation-defective MTAg, whereas mutations outside of this sequence allowed mutants to retain near-wild-type transformation capabilities. Transformation-defective mutants with mutations in the NPTY region behaved similarly to the mutant with the original Pro-248-to-Leu-248 mutation when assayed for associated proteins and activities in vitro; that is, they retained a full complement of wild-type activities and associated proteins. Further, insertion of the tetrameric sequence NPTY downstream of the mutated motif restored transforming abilities to these mutants. Thus, the tetrameric sequence NPTY in MTAg appears to represent a well-defined functional motif of MTAg.

Original languageEnglish (US)
Pages (from-to)5770-5776
Number of pages7
JournalJournal of Virology
Volume66
Issue number10
StatePublished - Oct 1992
Externally publishedYes

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Polyomavirus
Polyomavirus Transforming Antigens
Viral Tumor Antigens
antigens
mutants
Mutation
mutation
LDL Receptors
low density lipoprotein
receptors
Amino Acid Receptors
Amino Acids
Proteins
Insertional Mutagenesis
Amino Acid Substitution
amino acids
proteins
amino acid substitution
transposons
Phosphotransferases

ASJC Scopus subject areas

  • Immunology

Cite this

Druker, B., Sibert, L., & Roberts, T. M. (1992). Polyomavirus middle T-antigen NPTY mutants. Journal of Virology, 66(10), 5770-5776.

Polyomavirus middle T-antigen NPTY mutants. / Druker, Brian; Sibert, Lisa; Roberts, Tmomas M.

In: Journal of Virology, Vol. 66, No. 10, 10.1992, p. 5770-5776.

Research output: Contribution to journalArticle

Druker, B, Sibert, L & Roberts, TM 1992, 'Polyomavirus middle T-antigen NPTY mutants', Journal of Virology, vol. 66, no. 10, pp. 5770-5776.
Druker B, Sibert L, Roberts TM. Polyomavirus middle T-antigen NPTY mutants. Journal of Virology. 1992 Oct;66(10):5770-5776.
Druker, Brian ; Sibert, Lisa ; Roberts, Tmomas M. / Polyomavirus middle T-antigen NPTY mutants. In: Journal of Virology. 1992 ; Vol. 66, No. 10. pp. 5770-5776.
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