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PMID: 2523000 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Isolation and characterization of a viable adenovirus mutant defective in nuclear transport of the DNA-binding protein.

Journal of virology ·Vol. 63 ·No. 5 ·1989-05-00 ·Pages 2289-99

Cleghon V, Voelkerding K, Morin N, Delsert C, Klessig DF

Abstract

The isolation and characterization of an adenovirus mutant, Ad5dl802r1, containing two independent deletions in the 72-kilodalton (kDa) DNA-binding protein (DBP) gene is described. The two deletions remove amino acids 23 through 105 of DBP, resulting in the production of a 50-kDa product. Expression of this truncated DBP was delayed 12 to 24 h compared with that of the 72-kDa protein produced by wild-type adenovirus type 5. The DBP was located primarily in the cytoplasm of infected cells, whereas the wild-type product was predominantly nuclear. Therefore, DBP appears to contain a nuclear localization signal within the deleted region. Ad5dl802r1 DNA synthesis, viral late gene expression, and virus production were all delayed 12 to 24 h and were approximately 10-fold lower than with wild-type adenovirus type 5. These phenotypic properties can be accounted for by the delay in synthesis and the inefficient accumulation of the 50-kDa DBP within the nucleus of infected cells. The truncated DBP also lacks the majority of amino acids which are phosphorylated in the normal protein. The loss of these phosphorylation sites does not appear to seriously impair the ability of the protein to carry out its functions.

MeSH Terms
Adenovirus Early Proteins Adenoviruses, Human/genetics Base Sequence Biological Transport Cell Compartmentation Cell Nucleus/metabolism Chromosome Deletion DNA-Binding Proteins/genetics,metabolism Fluorescent Antibody Technique Gene Expression Regulation HeLa Cells Molecular Weight Mutation Nuclear Proteins/metabolism Oncogene Proteins, Viral/genetics,metabolism Phosphoproteins/metabolism Virus Replication
Chemicals
Adenovirus Early Proteins DNA-Binding Proteins Nuclear Proteins Oncogene Proteins, Viral Phosphoproteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Cleghon V
Waksman Institute of Microbiology Rutgers, State University of New Jersey, Piscataway 08855-0759.
Voelkerding K
Morin N
Delsert C
Klessig D F
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.63.5.2289-2299.1989
Published
1989-05-00
Pages
2289-99
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC250647
Subset
IM
Grants
PHS HHS · A123591 · United States
External Links
PubMed source DOI: 10.1128/JVI.63.5.2289-2299.1989 PMC Full Text View journal details
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