Cloning and characterization of a vertebrate cellular myosin regulatory light chain complementary DNA.
We have isolated two series of complementary DNAs (cDNAs) from a chicken gizzard cDNA library encoding two isoforms of phosphorylatable myosin regulatory light chain (RLC). One of the cDNAs encodes a previously isolated smooth muscle myosin RLC (also referred to as LC20-A); the other encodes a protein that shares 92% homology with the LC20-A isoform. The phosphorylatable threonine and serine residues at positions 18 and 19 of the two myosin RLC sequences are conserved. The two cDNAs are 81% homologous at the nucleotide level over the coding region; the 5' and 3' untranslated regions are divergent. Most of the DNA nonhomology in the coding region does not affect the protein sequence, indicating strong evolutionary conservation pressure to maintain the myosin RLC structure. Northern blot analysis using 3' untranslated region probes reveals restrictive tissue specific expression of one myosin RLC isoform (LC20-A) in smooth muscle tissue and not in other tissues examined. In contrast, the novel myosin RLC isoform messenger RNA (mRNA) is uniformly expressed in all smooth and nonmuscle tissues examined and is designated as cellular myosin RLC for this reason. Our results indicate that cellular and smooth muscle myosin RLC isoforms are distinct and are encoded by separate genes. This report describes the cloning of a novel vertebrate cellular myosin RLC mRNA that differs from previously characterized smooth muscle RLC isoform mRNAs in both primary sequence and expression pattern.
Zavodny
P J
PJ
Department of Biotechnology-Molecular Biology, Schering-Plough Corporation, Bloomfield, NJ 07003.
Petro
M E
ME
Lonial
H K
HK
Dailey
S H
SH
Narula
S K
SK
Leibowitz
P J
PJ
Kumar
C C
CC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004095:Dihydropyridines / Q000494:pharmacology*; D015870:Gene Expression; D005895:Gizzard, Avian / Q000737:chemistry; D008969:Molecular Sequence Data; D009219:Myosin-Light-Chain Kinase / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010766:Phosphorylation; D012333:RNA, Messenger / Q000032:analysis; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014018:Tissue Distribution
D004095:Dihydropyridines; D012333:RNA, Messenger; D004247:DNA; C065927:cilnidipine; D009219:Myosin-Light-Chain Kinase