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PMID: 1944267 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

A protein kinase-A recognition sequence is structurally linked to transformation by p59v-rel and cytoplasmic retention of p68c-rel.

Molecular and cellular biology ·Vol. 11 ·No. 12 ·1991-12-00 ·Pages 5867-77

Mosialos G, Hamer P, Capobianco AJ, Laursen RA, Gilmore TD

Abstract

The Rel family of proteins includes a number of proteins involved in transcriptional control, such as the retroviral oncoprotein v-Rel, c-Rel, the Drosophila melanogaster developmental protein Dorsal, and subunits of the transcription factor NF-kappa B. These proteins are related through a highly conserved domain of approximately 300 amino acids, called the Rel homology domain, that contains dimerization, DNA binding, and nuclear targeting functions. Also within the Rel homology domain, there is a conserved consensus sequence (Arg-Arg-Pro-Ser) for phosphorylation by cyclic AMP-dependent protein kinase (PKA). We used linker insertion mutagenesis and site-directed mutagenesis to determine the importance of this sequence for the transformation of avian spleen cells by v-Rel and the subcellular localization of c-Rel in chicken embryo fibroblasts (CEF). The insertion of 2 amino acids (Pro-Trp) within this sequence completely abolished transformation and transcriptional repression by v-Rel and resulted in a shift in the localization of c-Rel from cytoplasmic to nuclear in CEF. When the conserved Ser within the PKA recognition sequence was replaced by Ala, there was no significant effect on transformation and transcriptional repression by v-Rel or on cytoplasmic retention of c-Rel. However, when this Ser was changed to Asp or Glu, transformation and transcriptional repression by v-Rel were significantly inhibited and c-Rel showed a diffuse nuclear and cytoplasmic localization in CEF. Although a peptide containing the recognition sequence from v-Rel can be phosphorylated by PKA in vitro, this site is not constitutively phosphorylated to a high degree in vivo in transformed spleen cells incubated with okadaic acid. Our results indicate that the transforming and transcriptional repressing activities of v-Rel and the cytoplasmic retention of c-Rel are dependent on the structure of the conserved PKA recognition motif. In addition, they suggest that phosphorylation at the conserved PKA site could have a negative effect on transformation and transcriptional repression by v-Rel and induce the nuclear localization of c-Rel.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Blotting, Western Cell Transformation, Neoplastic Chickens Cytoplasm/metabolism DNA Molecular Sequence Data Mutation Oncogene Proteins v-rel Protein Kinases/metabolism Proto-Oncogene Proteins/genetics,metabolism Proto-Oncogene Proteins c-rel Retroviridae Proteins, Oncogenic/metabolism Substrate Specificity Transcription, Genetic Trypsin
Chemicals
Oncogene Proteins v-rel Proto-Oncogene Proteins Proto-Oncogene Proteins c-rel Retroviridae Proteins, Oncogenic DNA Protein Kinases Trypsin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Mosialos G
Department of Chemistry, Boston University, Massachusetts 02215.
Hamer P
Capobianco A J
Laursen R A
Gilmore T D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.12.5867-5877.1991
Published
1991-12-00
Pages
5867-77
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361736
Subset
IM
Grants
NCI NIH HHS · CA47763 · United States
External Links
PubMed source DOI: 10.1128/mcb.11.12.5867-5877.1991 PMC Full Text View journal details
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