Functional interaction of a novel cellular protein with the papillomavirus E2 transactivation domain

David E. Breiding, Francis Sverdrup, Martha J. Grossel, Nicola Moscufo, Waranya Boonchai, Elliot Androphy

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

The transactivation domain (AD) of bovine papillomavirus type 1 E2 stimulates gene expression and DNA replication. To identify cellular proteins that interact with this 215-amino-acid domain, we used a transactivation- defective mutant as bait in the yeast two-hybrid screen. In vitro and in vivo results demonstrate that the cDNA of one plasmid isolated in this screen encodes a 37-kDa nuclear protein that specifically binds to an 82-amino-acid segment within the E2 AD. Mutants with point mutations within this E2 domain were isolated based on their inability to interact with AMF-1 and were found to be unable to stimulate transcription. These mutants also exhibited defects in viral DNA replication yet retained binding to the viral E1 replication initiator protein. Overexpression of AMF-1 stimulated transactivation by both wild-type E2 and LexA fusion to the E2 AD, indicating that AMF-1 is a positive effector of the AD of E2. We conclude that interaction with AMF-1 is necessary for the transcriptional activation function of the E2 AD in mammalian cells.

Original languageEnglish (US)
Pages (from-to)7208-7219
Number of pages12
JournalMolecular and Cellular Biology
Volume17
Issue number12
StatePublished - Dec 1997
Externally publishedYes

Fingerprint

Transcriptional Activation
DNA Replication
Proteins
Bovine papillomavirus 1
Amino Acids
Viral DNA
Nuclear Proteins
Point Mutation
Plasmids
Complementary DNA
Yeasts
Gene Expression

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Breiding, D. E., Sverdrup, F., Grossel, M. J., Moscufo, N., Boonchai, W., & Androphy, E. (1997). Functional interaction of a novel cellular protein with the papillomavirus E2 transactivation domain. Molecular and Cellular Biology, 17(12), 7208-7219.

Functional interaction of a novel cellular protein with the papillomavirus E2 transactivation domain. / Breiding, David E.; Sverdrup, Francis; Grossel, Martha J.; Moscufo, Nicola; Boonchai, Waranya; Androphy, Elliot.

In: Molecular and Cellular Biology, Vol. 17, No. 12, 12.1997, p. 7208-7219.

Research output: Contribution to journalArticle

Breiding, DE, Sverdrup, F, Grossel, MJ, Moscufo, N, Boonchai, W & Androphy, E 1997, 'Functional interaction of a novel cellular protein with the papillomavirus E2 transactivation domain', Molecular and Cellular Biology, vol. 17, no. 12, pp. 7208-7219.
Breiding, David E. ; Sverdrup, Francis ; Grossel, Martha J. ; Moscufo, Nicola ; Boonchai, Waranya ; Androphy, Elliot. / Functional interaction of a novel cellular protein with the papillomavirus E2 transactivation domain. In: Molecular and Cellular Biology. 1997 ; Vol. 17, No. 12. pp. 7208-7219.
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