RRC ID 4282
Author Qadota H, McGaha LA, Mercer KB, Stark TJ, Ferrara TM, Benian GM.
Title A novel protein phosphatase is a binding partner for the protein kinase domains of UNC-89 (Obscurin) in Caenorhabditis elegans.
Journal Mol Biol Cell
Abstract Mutation of the Caenorhabditis elegans gene unc-89 results in disorganization of muscle A-bands. unc-89 encodes a giant polypeptide (900 kDa) containing two protein kinase domains, PK1 and PK2. Yeast two-hybrid screening using a portion of UNC-89 including PK2, yielded SCPL-1 (small CTD phosphatase-like-1), which contains a C terminal domain (CTD) phosphatase type domain. In addition to the PK2 domain, interaction with SCPL-1 required the putative autoinhibitory sequence, and immunoglobulin (Ig) and fibronectin type 3 (Fn3) domains lying N-terminal of the kinase domain. SCPL-1 also interacts with PK1, and it similarly requires the kinase domain and upstream Fn3 and Ig domains. Analogous regions from the two other giant kinases of C. elegans, twitchin and TTN-1, failed to interact with SCPL-1. The interaction between SCPL-1 and either Ig-Fn3-PK2 or Fn3-Ig-PK1 was confirmed by biochemical methods. The scpl-1b promoter is expressed in the same set of muscles as unc-89. Antibodies to SCPL-1 localize to the M-line and a portion of the I-band. Bacterially expressed SCPL-1 proteins have phosphatase activity in vitro with properties similar to previously characterized members of the CTD phosphatase family. RNA interference knockdown results in a defect in the function of egg-laying muscles. These studies suggest a new role for the CTD phosphatase family, that is, in muscle giant kinase signaling.
Volume 19(6)
Pages 2424-32
Published 2008-6-1
DOI 10.1091/mbc.e08-01-0053
PII E08-01-0053
PMID 18337465
PMC PMC2397297
MeSH Animals Caenorhabditis elegans / enzymology* Caenorhabditis elegans Proteins / chemistry* Caenorhabditis elegans Proteins / genetics Caenorhabditis elegans Proteins / metabolism* Gene Expression Regulation Muscle Proteins / chemistry* Muscle Proteins / metabolism* Oviposition Phosphoprotein Phosphatases / genetics Phosphoprotein Phosphatases / metabolism* Promoter Regions, Genetic / genetics Protein Binding Protein Kinases Protein Structure, Tertiary Reproducibility of Results Substrate Specificity Two-Hybrid System Techniques
IF 3.791
Times Cited 36
WOS Category CELL BIOLOGY
Resource
C.elegans tm752