Identification of casein kinase I substrates by in vitro expression cloning screening.
Journal Article (Journal Article)
Casein kinase I (CKI) is a widely expressed protein kinase family implicated in diverse processes including membrane trafficking, DNA repair, and circadian rhythm. Despite the large number of CKI genes, few biologically relevant substrates have been identified. As an approach to better defining the spectrum of CKI substrates, we extended a recently described in vitro expression cloning (IVEC) strategy. Polypeptides pools were screened for kinase-dependent electrophoretic mobility shifts. Ten putative CKI substrates were isolated from an initial sample of 3000 random cDNA clones. Candidate substrates include proteins involved in RNA metabolism (a putative RNA helicase, the nucleolar protein hNOP56, and hnRNP A1, and ribosomal proteins L4, L8, and L13), as well as keratin 17, a necdin-related protein, and the calcium-binding proteins desmoglein 2 and annexin II. The same pools were also screened with active ERK2, and four substrates identified: aldolase, NSD-like protein, uracil-DNA glycosylase, and HHR23A. IVEC is an effective method to identify novel protein kinase substrates.
Full Text
Duke Authors
Cited Authors
- Gao, ZH; Metherall, J; Virshup, DM
Published Date
- February 16, 2000
Published In
Volume / Issue
- 268 / 2
Start / End Page
- 562 - 566
PubMed ID
- 10679243
International Standard Serial Number (ISSN)
- 0006-291X
Digital Object Identifier (DOI)
- 10.1006/bbrc.2000.2168
Language
- eng
Conference Location
- United States