Isolation and characterization of pmk-(1-3): three p38 homologs in Caenorhabditis elegans

Mol Cell Biol Res Commun. 2001 Nov;4(6):337-44. doi: 10.1006/mcbr.2001.0300.

Abstract

p38, a member of the mitogen-activated protein kinase (MAPK) superfamily, is activated in response to a variety of cellular stresses and ligands. Since the genome of the nematode C. elegans has been sequenced, we sought to identify and characterize the nematode homolog of mammalian p38. By sequence analysis and RT-PCR, we isolated cDNAs encoding three kinases, PMK-1, PMK-2, and PMK-3, which we call p38 map kinases due to their high sequence identity with p38. The three genes are contiguous on chromosome IV and comprise an operon. By use of a GFP reporter, we found that the promoter of the pmks is active throughout the intestine. An active form of MAPK/ERK kinase 6 (MEK6) phosphorylated and activated recombinant PMK-1 and PMK-2 in vitro. PMK-1 and PMK-2 phosphorylated activating transcription factor-2 (ATF-2), indicating an activity similar to mammalian p38. When transfected into mammalian cells, these kinases, like p38, are stimulated by osmotic stresses.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Activating Transcription Factor 2
  • Amino Acid Sequence
  • Animals
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans Proteins*
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Cell Line
  • Cloning, Molecular
  • Cyclic AMP Response Element-Binding Protein / metabolism
  • DNA, Complementary / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Fungal Proteins*
  • Green Fluorescent Proteins
  • Humans
  • Intestinal Mucosa / metabolism
  • Luminescent Proteins / metabolism
  • MAP Kinase Kinase 6
  • Mitogen-Activated Protein Kinases / chemistry*
  • Mitogen-Activated Protein Kinases / genetics*
  • Mitogen-Activated Protein Kinases / isolation & purification
  • Mitogen-Activated Protein Kinases / metabolism
  • Molecular Sequence Data
  • Osmosis
  • Phenotype
  • Phosphorylation
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA / metabolism
  • RNA Splicing
  • Recombinant Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factors / metabolism
  • Transfection
  • p38 Mitogen-Activated Protein Kinases

Substances

  • ATF2 protein, human
  • Activating Transcription Factor 2
  • Caenorhabditis elegans Proteins
  • Cyclic AMP Response Element-Binding Protein
  • DNA, Complementary
  • Fungal Proteins
  • Luminescent Proteins
  • Recombinant Proteins
  • Transcription Factors
  • Green Fluorescent Proteins
  • RNA
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Mitogen-Activated Protein Kinases
  • Pmk1 protein, Magnaporthe grisea
  • p38 Mitogen-Activated Protein Kinases
  • pmk-2 protein, C elegans
  • pmk-3 protein, C elegans
  • MAP Kinase Kinase 6
  • MAP2K6 protein, human