Cloning of rat MEK kinase 1 cDNA reveals an endogenous membrane-associated 195-kDa protein with a large regulatory domain.
The coding sequence of rat MEK kinase 1 (MEKK1) has been determined from multiple, independent cDNA clones. The cDNA is full-length based on the presence of stop codons in all three reading frames of the 5' untranslated region. Probes from the 5' and the 3' coding sequences both hybridize to a 7-kb mRNA. The open reading frame is 4.5 kb and predicts a protein with molecular mass of 161,225 Da, which is twice the size of the previously published MEKK1 sequence and reveals 801 amino acids of novel coding sequence. The novel sequence contains two putative pH domains, two proline-rich regions, and a cysteine-rich region. Antisera to peptides derived from this new sequence recognize an endogenous protein in human and rodent cells of 195 kDa, consistent with the size of the expressed rat MEKK1 clone. Endogenous and recombinant rat MEKK1 are enriched in membranes; little of either is found in soluble fractions. Expression of recombinant rat MEKK1 leads to activation of three mitogen-activated protein kinase modules in the order c-Jun N-terminal kinase/stress-activated protein kinase > p38 mitogen-activated protein kinase = extracellular signal-regulated kinase 2.
Xu
S
S
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, 75235-9041, USA.
Robbins
D J
DJ
Christerson
L B
LB
English
J M
JM
Vanderbilt
C A
CA
Cobb
M H
MH
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary; D015723:Gene Library; D006801:Humans; D008099:Liver / Q000201:enzymology; D048728:MAP Kinase Kinase Kinase 1; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D016716:PC12 Cells; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis* / Q000737:chemistry; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000737:chemistry; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D015183:Restriction Mapping; D014162:Transfection
D018076:DNA, Complementary; D011994:Recombinant Proteins; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D048728:MAP Kinase Kinase Kinase 1; C482576:MAP3K1 protein, human