Dissection of functional domains in the adenovirus 2 early 1b 55k polypeptide by suppressor-linker insertional mutagenesis

Patricia R Yew, C. Cheng Kao, Arnold J. Berk

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

To determine whether the viral replication functions of the adenovirus E1 B 55K protein play a role in its ability to transform cloned rat embryo fibroblast cells in culture, we constructed an extensive series of insertion mutations throughout the 55K gene. The mutations were recombined into infectious virus and characterized for their abilities to produce stable 55K protein in HeLa cells, replicate virus in HeLa cells, express late viral proteins efficiently, and transform CREF cells following infection. Mutant 55K transforming activity in primary baby rat kidney cells was also assayed following DNA transfection. The functions required for viral replication are encoded in several patches of the 55K linear sequence, while the CREF transforming functions are sensitive to all of the insertions. An insertion at amino acid 380 created a mutant virus which was reduced in transforming activity, but was not reduced for viral replication. Therefore, a function required for efficient transformation of CREF cells can be separated from functions required for late gene expression and viral replication. Transformation of BRK cells following DNA transfection was reduced by complete disruption of the 55K protein gene, but was not significantly affected by any of the insertions.

Original languageEnglish (US)
Pages (from-to)795-805
Number of pages11
JournalVirology
Volume179
Issue number2
DOIs
StatePublished - 1990
Externally publishedYes

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Insertional Mutagenesis
Adenoviridae
Dissection
Peptides
Viruses
HeLa Cells
Transfection
DNA
Viral Proteins
Proteins
Embryonic Structures
Cell Culture Techniques
Fibroblasts
Kidney
Gene Expression
Amino Acids
Mutation
Infection
Genes

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Dissection of functional domains in the adenovirus 2 early 1b 55k polypeptide by suppressor-linker insertional mutagenesis. / Yew, Patricia R; Cheng Kao, C.; Berk, Arnold J.

In: Virology, Vol. 179, No. 2, 1990, p. 795-805.

Research output: Contribution to journalArticle

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