Structure and expression of a calcium-binding protein gene contained within a calmodulin-regulated protein kinase gene.
We have determined the first genomic structure and characterized the mRNA and protein products of a novel vertebrate gene that encodes a calcium-binding protein with amino acid sequence identity to a protein kinase domain. The elucidation of the complete DNA sequence of this transcription unit and adjacent genomic DNA, Southern blot and polymerase chain reaction analyses of cellular genomic DNA, and examination of mRNA and protein species revealed that the calcium-binding kinase-related protein (KRP)-encoding gene is contained within the gene for a calmodulin-regulated protein kinase, myosin light-chain kinase (MLCK). The KRP gene transcription unit is composed of three exons and a 5'-flanking sequence containing a canonical TATA box motif. The TATA box, the transcription initiation site, and the first 109 nucleotides of the 5' noncoding region of the KRP mRNA correspond to an MLCK gene intron sequence. Both KRP and MLCK are produced in the same adult chicken tissue in relatively high abundance from a single contiguous stretch of genomic DNA and utilize the same reading frame and common exons to produce distinct mRNAs (2.7 and 5.5 kb, respectively) that encode proteins with dissimilar biochemical functions. There appears to be no precedent in vertebrate molecular biology for such a relationship. This may represent a mechanism whereby functional diversity can be achieved within the same vertebrate tissue by use of common exons to produce shuffled domains with identical amino acid sequences in different molecular contexts.
Collinge
M
M
Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232-6600.
Matrisian
P E
PE
Zimmer
W E
WE
Shattuck
R L
RL
Lukas
T J
TJ
Van Eldik
L J
LJ
Watterson
D M
DM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification; D002147:Calmodulin / Q000494:pharmacology; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D005796:Genes; D016678:Genome; D005895:Gizzard, Avian / Q000201:enzymology; D015151:Immunoblotting; D007438:Introns; D016547:Kinesins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009124:Muscle Proteins / Q000235:genetics* / Q000302:isolation & purification; D009130:Muscle, Smooth / Q000201:enzymology*; D009219:Myosin-Light-Chain Kinase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D016385:TATA Box
D002135:Calcium-Binding Proteins; D002147:Calmodulin; D009124:Muscle Proteins; D012333:RNA, Messenger; C074152:kinesin-II; D012313:RNA; D004247:DNA; D009219:Myosin-Light-Chain Kinase; D016547:Kinesins