Cloning and in Situ Hybridization of Type 2A and 2B Rat Skeletal Muscle Myosin Tail Region: Implications for Filament Assembly

R. L. Lieber, S. C. Bodine, T. J. Burkholder, D. J. Pierotti, A. F. Ryan

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Changes in fast myosin expression play a critical role in skeletal muscle adaptation. Two fast myosin isoforms, type 2A and type 2B, are commonly expressed by fast muscle fibers but their sequences have not been determined to allow mRNA expression studies. A complete set of rat skeletal muscle myosins was amplified by PCR of cDNAs derived from skeletal muscle mRNA, cloned in a TA cloning vector, and sequenced. Specificity was demonstrated by in situ hybridization against skeletal muscle and myosin protein identification using monoclonal antibodies. Two novel sequences were cloned: A type 2A myosin which consisted of a 642 bp segment from the 3′ end and a type 2B myosin which consisted of a 624 bp segment also from the 3′ end. This region encodes that portion of the myosin molecule implicated in the control of filament assembly. The two fast myosins showed 88% homology in the open reading frame and 95% homology at the amino acid level. Based on this homology, it is unlikely that selective myosin filament assembly occurs during muscle fiber type transformation between type 2A and 2B.

Original languageEnglish (US)
Pages (from-to)1312-1318
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume197
Issue number3
DOIs
StatePublished - Dec 30 1993
Externally publishedYes

Fingerprint

Skeletal Muscle Myosins
Cloning
Myosins
In Situ Hybridization
Tail
Rats
Organism Cloning
Muscle
Skeletal Muscle
Muscles
Genetic Vectors
Messenger RNA
Muscle Proteins
Fibers
Open Reading Frames
Protein Isoforms
Complementary DNA
Monoclonal Antibodies
Amino Acids
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

Cloning and in Situ Hybridization of Type 2A and 2B Rat Skeletal Muscle Myosin Tail Region : Implications for Filament Assembly. / Lieber, R. L.; Bodine, S. C.; Burkholder, T. J.; Pierotti, D. J.; Ryan, A. F.

In: Biochemical and Biophysical Research Communications, Vol. 197, No. 3, 30.12.1993, p. 1312-1318.

Research output: Contribution to journalArticle

Lieber, R. L. ; Bodine, S. C. ; Burkholder, T. J. ; Pierotti, D. J. ; Ryan, A. F. / Cloning and in Situ Hybridization of Type 2A and 2B Rat Skeletal Muscle Myosin Tail Region : Implications for Filament Assembly. In: Biochemical and Biophysical Research Communications. 1993 ; Vol. 197, No. 3. pp. 1312-1318.
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