Nebulin interacts with CapZ and regulates thin filament architecture within the Z-disc

Christopher T. Pappas, Nandini Bhattacharya, John A. Cooper, Carol Gregorio

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

The barbed ends of actin filaments in striated muscle are anchored within the Z-disc and capped by CapZ; this protein blocks actin polymerization and depolymerization in vitro. The mature lengths of the thin filaments are likely specified by the giant "molecular ruler" nebulin, which spans the length of the thin filament. Here, we report that CapZ specifically interacts with the C terminus of nebulin (modules 160-164) in blot overlay, solid-phase binding, tryptophan fluorescence, and SPOTs membrane assays. Binding of nebulin modules 160-164 to CapZ does not affect the ability of CapZ to cap actin filaments in vitro, consistent with our observation that neither of the two C-terminal actin binding regions of CapZ is necessary for its interaction with nebulin. Knockdown of nebulin in chick skeletal myotubes using small interfering RNA results in a reduction of assembled CapZ, and, strikingly, a loss of the uniform alignment of the barbed ends of the actin filaments. These data suggest that nebulin restricts the position of thin filament barbed ends to the Z-disc via a direct interaction with CapZ. We propose a novel molecular model of Z-disc architecture in which nebulin interacts with CapZ from a thin filament of an adjacent sarcomere, thus providing a structural link between sarcomeres.

Original languageEnglish (US)
Pages (from-to)1837-1847
Number of pages11
JournalMolecular Biology of the Cell
Volume19
Issue number5
DOIs
StatePublished - May 2008

Fingerprint

Actin Cytoskeleton
Sarcomeres
CapZ Actin Capping Protein
Actins
Molecular Models
Striated Muscle
Skeletal Muscle Fibers
nebulin
Tryptophan
Polymerization
Small Interfering RNA
Fluorescence
Membranes
In Vitro Techniques

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Nebulin interacts with CapZ and regulates thin filament architecture within the Z-disc. / Pappas, Christopher T.; Bhattacharya, Nandini; Cooper, John A.; Gregorio, Carol.

In: Molecular Biology of the Cell, Vol. 19, No. 5, 05.2008, p. 1837-1847.

Research output: Contribution to journalArticle

Pappas, Christopher T. ; Bhattacharya, Nandini ; Cooper, John A. ; Gregorio, Carol. / Nebulin interacts with CapZ and regulates thin filament architecture within the Z-disc. In: Molecular Biology of the Cell. 2008 ; Vol. 19, No. 5. pp. 1837-1847.
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