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Molecular cloning and characterization of a novel protein kinase C-interacting protein with structural motifs related to RBCC family proteins.
- Source :
-
Biochemical and biophysical research communications [Biochem Biophys Res Commun] 1998 Mar 17; Vol. 244 (2), pp. 353-9. - Publication Year :
- 1998
-
Abstract
- A novel protein kinase C (PKC)-interacting protein was identified by the yeast two-hybrid screening using the regulatory domain of PKC beta I as a bait. The protein contained several structural motifs such as two putative coiled-coil regions, a RING-finger, a B-box, and a B-box-like motif in the order from NH2- to COOH-terminals. The molecular organization of the protein resembles the structure of the RBCC protein family proteins which usually have a RING-finger, a B-box, and a coiled-coil region. Therefore, the protein identified was designated as RBCK1 (RBCC protein interacting with PKC 1). Northern blot analysis showed that RBCK1 gene is expressed ubiquitously among rat tissues. RBCK1 protein associated with PKC beta I and PKC zeta when coexpressed in cultured mammalian cells. By the polymerase chain reaction-assisted DNA-binding site selection and the electrophoretic mobility shift assay, RBCK1 protein was shown to bind to several DNA fragments containing TGG-rich sequences. When the yeast GAL4 DNA-binding domain fused RBCK1 protein was expressed in COS-7 cells harboring the luciferase gene placed under a synthetic promoter containing GAL4-binding sites, the fusion protein showed enhanced transcriptional activity comparing with the GAL4 DNA-binding domain. These results suggest that RBCK1 protein might be a transcription factor that has a role in the signaling pathway through PKC.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Binding Sites genetics
Cloning, Molecular
DNA Primers genetics
DNA, Complementary genetics
Molecular Sequence Data
Nerve Tissue Proteins metabolism
Polymerase Chain Reaction
Protein Kinase C beta
Rats
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae metabolism
Signal Transduction
Transcription Factors chemistry
Transcription Factors genetics
Transcription Factors metabolism
Transcriptional Activation
Isoenzymes metabolism
Nerve Tissue Proteins chemistry
Nerve Tissue Proteins genetics
Protein Kinase C metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0006-291X
- Volume :
- 244
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Biochemical and biophysical research communications
- Publication Type :
- Academic Journal
- Accession number :
- 9514928
- Full Text :
- https://doi.org/10.1006/bbrc.1998.8270