Cloning of rat MEK kinase 1 cDNA reveals an endogenous membrane-associated 195-kDa protein with a large regulatory domain

Shuichan Xu, David J Robbins, Lori B. Christerson, Jessie M. English, Colleen A. Vanderbilt, Melanie H. Cobb

Research output: Contribution to journalArticle

121 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)5291-5295
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number11
DOIs
StatePublished - May 28 1996
Externally publishedYes

Fingerprint

MAP Kinase Kinase Kinase 1
Organism Cloning
Complementary DNA
Membranes
Proteins
Clone Cells
Reading Frames
Terminator Codon
JNK Mitogen-Activated Protein Kinases
5' Untranslated Regions
Mitogen-Activated Protein Kinase 1
p38 Mitogen-Activated Protein Kinases
Heat-Shock Proteins
Mitogen-Activated Protein Kinases
Proline
Protein Kinases
Open Reading Frames
Cysteine
Immune Sera
Rodentia

Keywords

  • cysteine-rich region
  • membrane-associated protein kinase
  • mitogen-activated protein kinase pathways
  • pH domain
  • proline-rich region

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Cloning of rat MEK kinase 1 cDNA reveals an endogenous membrane-associated 195-kDa protein with a large regulatory domain. / Xu, Shuichan; Robbins, David J; Christerson, Lori B.; English, Jessie M.; Vanderbilt, Colleen A.; Cobb, Melanie H.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, No. 11, 28.05.1996, p. 5291-5295.

Research output: Contribution to journalArticle

Xu, Shuichan ; Robbins, David J ; Christerson, Lori B. ; English, Jessie M. ; Vanderbilt, Colleen A. ; Cobb, Melanie H. / Cloning of rat MEK kinase 1 cDNA reveals an endogenous membrane-associated 195-kDa protein with a large regulatory domain. In: Proceedings of the National Academy of Sciences of the United States of America. 1996 ; Vol. 93, No. 11. pp. 5291-5295.
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