A LIM-9 (FHL) / SCPL-1 (SCP) Complex Interacts with the C-terminal Protein Kinase Regions of UNC-89 (Obscurin) in Caenorhabditis elegans Muscle

Ge Xiong, Hiroshi Qadota, Kristina B. Mercer, Lee Anne McGaha, Andres F. Oberhauser, Guy M. Benian

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The C. elegans gene unc-89 encodes a set of mostly giant polypeptides (up to 900 kDa) that contain multiple immunoglobulin (Ig) and fibronectin type 3 (Fn3), a triplet of SH3-DH-PH, and two protein kinase domains. The loss of function mutant phenotype and localization of antibodies to UNC-89 proteins indicate that the function of UNC-89 is to help organize sarcomeric A-bands, especially M-lines. Recently, we reported that each of the protein kinase domains interacts with SCPL-1, which contains a CTD-type protein phosphatase domain. Here, we report that SCPL-1 interacts with LIM-9 (FHL), a protein that we first discovered as an interactor of UNC-97 (PINCH) and UNC-96, components of an M-line costamere in nematode muscle. We show that LIM-9 can interact with UNC-89 through its first kinase domain and a portion of unique sequence lying between the two kinase domains. All the interactions were confirmed by biochemical methods. A yeast three-hybrid assay demonstrates a ternary complex between the two protein kinase regions and SCPL-1. Evidence that the UNC-89/SCPL-1 interaction occurs in vivo was provided by showing that over-expression of SCPL-1 results in disorganization of UNC-89 at M-lines. We suggest two structural models for the interactions of SCPL-1 and LIM-9 with UNC-89 at the M-line.

Original languageEnglish (US)
Pages (from-to)976-988
Number of pages13
JournalJournal of Molecular Biology
Volume386
Issue number4
DOIs
StatePublished - Mar 6 2009
Externally publishedYes

Fingerprint

Caenorhabditis elegans
Protein Kinases
Protein Kinase C
Muscles
Costameres
Phosphotransferases
Myeloma Proteins
Two-Hybrid System Techniques
Phosphoprotein Phosphatases
Structural Models
Fibronectins
Immunoglobulins
Proteins
Phenotype
Peptides
Antibodies
Genes
Protein Domains

Keywords

  • C. elegans
  • CTD phosphatase
  • FHL
  • muscle
  • obscurin

ASJC Scopus subject areas

  • Molecular Biology

Cite this

A LIM-9 (FHL) / SCPL-1 (SCP) Complex Interacts with the C-terminal Protein Kinase Regions of UNC-89 (Obscurin) in Caenorhabditis elegans Muscle. / Xiong, Ge; Qadota, Hiroshi; Mercer, Kristina B.; McGaha, Lee Anne; Oberhauser, Andres F.; Benian, Guy M.

In: Journal of Molecular Biology, Vol. 386, No. 4, 06.03.2009, p. 976-988.

Research output: Contribution to journalArticle

Xiong, Ge ; Qadota, Hiroshi ; Mercer, Kristina B. ; McGaha, Lee Anne ; Oberhauser, Andres F. ; Benian, Guy M. / A LIM-9 (FHL) / SCPL-1 (SCP) Complex Interacts with the C-terminal Protein Kinase Regions of UNC-89 (Obscurin) in Caenorhabditis elegans Muscle. In: Journal of Molecular Biology. 2009 ; Vol. 386, No. 4. pp. 976-988.
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