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PMID: 2192265 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Induction of transformation and DNA synthesis after microinjection of raf proteins.

Molecular and cellular biology ·Vol. 10 ·No. 7 ·1990-07-00 ·Pages 3828-33

Smith MR, Heidecker G, Rapp UR, Kung HF

Abstract

Full-length and N-terminal deletion mutants of human c-raf-1 cDNA were cloned into Escherichia coli expression plasmids. Bacterially expressed c-raf proteins were purified by anion-exchange, gel filtration, and affinity chromatography. Microinjection of mutant c-raf proteins into G0-arrested NIH 3T3 cells induced DNA synthesis and morphological transformation, whereas microinjection of full-length c-raf had no effect. The amino terminus of the raf protein has an important negative regulatory influence; alteration of this region resulted in increased kinase activity and oncogenicity.

MeSH Terms
Animals Cell Transformation, Neoplastic Cells, Cultured Cloning, Molecular DNA Replication Escherichia coli/genetics Humans Mice Microinjections Mutation Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins/administration & dosage,genetics,metabolism Proto-Oncogene Proteins c-raf Recombinant Proteins/administration & dosage,metabolism Restriction Mapping
Chemicals
Proto-Oncogene Proteins Recombinant Proteins Protein-Tyrosine Kinases Proto-Oncogene Proteins c-raf
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Smith M R
Biological Carcinogenesis and Development Program, Program Resources, Inc., National Cancer Institute-Frederick Cancer Research Facility, Maryland 21701.
Heidecker G
Rapp U R
Kung H F
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26 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.7.3828-3833.1990
Published
1990-07-00
Pages
3828-33
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360849
Subset
IM
Grants
NCI NIH HHS · N01-CO-74102 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.7.3828-3833.1990 PMC Full Text View journal details
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