PMID- 10198165
OWN - NLM
STAT- MEDLINE
DCOM- 19990506
LR  - 20190905
IS  - 0888-7543 (Print)
IS  - 0888-7543 (Linking)
VI  - 57
IP  - 2
DP  - 1999 Apr 15
TI  - A single human myosin light chain kinase gene (MLCK; MYLK).
PG  - 256-67
AB  - The myosin light chain kinase (MLCK) gene, a muscle member of the immunoglobulin 
      gene superfamily, yields both smooth muscle and nonmuscle cell isoforms. Both
      isoforms are known to regulate contractile activity via
      calcium/calmodulin-dependent myosin light chain phosphorylation. We previously
      cloned from a human endothelial cell (EC) cDNA library a high-molecular-weight
      nonmuscle MLCK isoform (EC MLCK (MLCK 1) with an open reading frame that encodes 
      a protein of 1914 amino acids. We now describe four novel nonmuscle MLCK isoforms
      (MLCK 2, 3a, 3b, and 4) that are the alternatively spliced variants of an mRNA
      precursor that is transcribed from a single human MLCK gene. The primary
      structure of the cDNA encoding the nonmuscle MLCK isoform 2 is identical to the
      previously published human nonmuscle MLCK (MLCK 1) (J. G. N. Garcia et al., 1997,
      Am. J. Respir. Cell Mol. Biol. 16, 489-494) except for a deletion of nucleotides 
      1428-1634 (D2). The full nucleotide sequence of MLCK isoforms 3a and 3b and
      partial sequence for MLCK isoform 4 revealed identity to MLCK 1 except for
      deletions at nucleotides 5081-5233 (MLCK 3a, D3), double deletions of nucleotides
      1428-1634 and 5081-5233 (MLCK 3b), and nucleotide deletions 4534-4737 (MLCK 4,
      D4). Northern blot analysis demonstrated the extended expression pattern of the
      nonmuscle MLCK isoform(s) in both human adult and human fetal tissues. RT-PCR
      using primer pairs that were designed to detect specifically nonmuscle MLCK
      isoforms 2, 3, and 4 deletions (D2, D3, and D4) confirmed expression in both
      human adult and human fetal tissues (lung, liver, brain, and kidney) and in human
      endothelial cells (umbilical vein and dermal). Furthermore, relative quantitative
      expression studies demonstrated that the nonmuscle MLCK isoform 2 is the dominant
      splice variant expressed in human tissues and cells. Further analysis of the
      human MLCK gene revealed that the MLCK 2 isoform represents the deletion of an
      independent exon flanked by 5' and 3' neighboring introns of 0.6 and 7.0 kb,
      respectively. Together these studies demonstrate for the first time that the
      human MLCK gene yields multiple nonmuscle MLCK isoforms by alternative splicing
      of its transcribed mRNA precursor with differential distribution of these
      isoforms in various human tissues and cells.
CI  - Copyright 1999 Academic Press.
FAU - Lazar, V
AU  - Lazar V
AD  - Division of Pulmonary and Critical Care Medicine, Johns Hopkins University School
      of Medicine, Baltimore, Maryland 21224, USA.
FAU - Garcia, J G
AU  - Garcia JG
LA  - eng
GR  - HL 50533/HL/NHLBI NIH HHS/United States
GR  - HL 58064/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Genomics
JT  - Genomics
JID - 8800135
RN  - 0 (Isoenzymes)
RN  - 0 (RNA, Messenger)
RN  - EC 2.7.11.18 (Myosin-Light-Chain Kinase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Blotting, Northern
MH  - Cell Line
MH  - Cloning, Molecular
MH  - Endothelium, Vascular/cytology/metabolism
MH  - Exons
MH  - Gene Deletion
MH  - Gene Expression Regulation
MH  - Humans
MH  - Isoenzymes/*genetics
MH  - Molecular Sequence Data
MH  - Myosin-Light-Chain Kinase/*genetics
MH  - RNA Splicing
MH  - RNA, Messenger/genetics/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Tissue Distribution
MH  - Transcription, Genetic
EDAT- 1999/04/13 00:00
MHDA- 1999/04/13 00:01
CRDT- 1999/04/13 00:00
PHST- 1999/04/13 00:00 [pubmed]
PHST- 1999/04/13 00:01 [medline]
PHST- 1999/04/13 00:00 [entrez]
AID - S0888754399957749 [pii]
AID - 10.1006/geno.1999.5774 [doi]
PST - ppublish
SO  - Genomics. 1999 Apr 15;57(2):256-67. doi: 10.1006/geno.1999.5774.