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PMID: 8497045 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Human papillomavirus type 18 E7 protein requires intact Cys-X-X-Cys motifs for zinc binding, dimerization, and transformation but not for Rb binding.

Journal of virology ·Vol. 67 ·No. 6 ·1993-06-00 ·Pages 3142-50

McIntyre MC, Frattini MG, Grossman SR, Laimins LA

Abstract

Human papillomavirus type 18 (HPV-18) E7 proteins bind zinc through Cys-X-X-Cys repeats located at the C terminus of the protein. In order to examine the role of these cysteine motifs in E7 function, we expressed the HPV-18 E7 protein in bacteria and found that purified E7 forms a dimer through interactions with zinc. Mutants with single mutations within the Cys-X-X-Cys motifs bound a reduced level of zinc in a zinc blot assay, while a double mutant lost all zinc-binding activity. When expressed in vivo, none of the mutants cooperated with an activated ras oncogene to transform primary rat embryo fibroblasts, but all mutants retained nearly wild-type Rb-binding activity. The results indicate that the cysteine motifs play an important role in transformation by HPV-18 E7 but do not contribute to Rb binding.

MeSH Terms
Adenovirus E1A Proteins/genetics Amino Acid Sequence Animals Cell Transformation, Viral Cells, Cultured Conserved Sequence Cysteine/metabolism DNA Mutational Analysis DNA-Binding Proteins Edetic Acid/pharmacology Escherichia coli/genetics Half-Life Models, Chemical Molecular Sequence Data Oncogene Proteins, Viral/genetics,isolation & purification,metabolism,pharmacokinetics Protein Conformation/drug effects Rats Recombinant Proteins/metabolism Repetitive Sequences, Nucleic Acid Retinoblastoma Protein/metabolism Rubidium/metabolism Zinc/metabolism
Chemicals
Adenovirus E1A Proteins DNA-Binding Proteins E7 protein, Human papillomavirus type 18 Oncogene Proteins, Viral Recombinant Proteins Retinoblastoma Protein Edetic Acid Zinc Cysteine Rubidium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
McIntyre M C
Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637.
Frattini M G
Grossman S R
Laimins L A
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.6.3142-3150.1993
Published
1993-06-00
Pages
3142-50
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237652
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.67.6.3142-3150.1993 PMC Full Text View journal details
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