The two essential light chains of carp fast skeletal myosin, LC1 and LC3, are encoded by distinct genes and change their molar ratio following temperature acclimation.
cDNA libraries were constructed from fast skeletal muscles of carp acclimated to 10 and 30 degrees C for a minimum of 5 weeks and were screened for myosin alkali light chains, LC1 and LC3, using an anti-skipjack LC1 polyclonal antibody. Two types of LC1 cDNA clone were isolated and termed LC1a and LC1b: their nucleotide sequences showed 92% homology. The ratio of LC1a to LC1b cDNA clones isolated was approximately 3:1, showing no apparent changes following temperature acclimation. The occurrence of the two isoforms was further confirmed by N-terminal amino acid sequencing of purified LC1. No isoform was, however, detected for LC3, while homology in the overlapping region between LC1a and LC3 cDNAs was only 65% even after the most probable alignment. Southern blot analyses probed with cDNA clones specific to LC1a and LC3 showed different hybridization patterns from each other, demonstrating that carp LC1 and LC3 are encoded by different genes. These results are in marked contrast to those from higher vertebrates which express LC1 and LC3 from a single gene by alternative RNA transcription and two modes of splicing. Northern blot analysis showed that the ratios of LC3/LC1 mRNAs were significantly higher (3.93) in 30 degrees C-acclimated than in 10 degrees C-acclimated (3.10) carp.
Hirayama
Y
Y
Laboratory of Aquatic Molecular Biology and Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Kanoh
S
S
Nakaya
M
M
Watabe
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D005786:Gene Expression Regulation; D005796:Genes; D008969:Molecular Sequence Data; D018994:Myosin Light Chains / Q000235:genetics*; D010449:Peptide Mapping; D015183:Restriction Mapping; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D018076:DNA, Complementary; D018994:Myosin Light Chains