TY - JOUR
T1 - Cloning and in situ hybridization of type 2A and 2B rat skeletal muscle myosin tail region
T2 - Implications for filament assembly
AU - Lieber, Richard L.
AU - Bodine, Sue C.
AU - Burkholder, Thomas J.
AU - Pierotti, David J.
AU - Ryan, Allen F.
PY - 1993/12/30
Y1 - 1993/12/30
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0027762693&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027762693&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1993.2620
DO - 10.1006/bbrc.1993.2620
M3 - Article
C2 - 8280148
AN - SCOPUS:0027762693
SN - 0006-291X
VL - 197
SP - 1312
EP - 1318
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -