Molecular Cloning and Expression Determination of p38 MAPK from the Liver and Kidney of Silver Carp

J Biochem Mol Toxicol. 2016 May;30(5):224-31. doi: 10.1002/jbt.21781. Epub 2016 Feb 11.

Abstract

The sequence of p38MAPK in silver carp (Hm-p38a) was cloned and sequenced. Additionally, the acute toxicity of crude microcystins (MCs) on silver carp and induction expression of Hm-p38a by MCs exposure were also determined in this study. The results reveal that the length of Hm-p38a is 2418 bp and it contains a 1086 bp open reading frame. Hm-p38a could encode 361 amino acids. Sequence analysis indicates that Hm-p38a contains the conserved structures of Thr-Gly-Tyr motif and substrate binding site Ala-Thr-Arg-Trp, and it is highly conserved in fish. Phylogenetic analysis reveals that Hm-p38a is more closely related to fish than mammals and it belongs to p38α subfamily. Moreover, Hm-p38a in silver carp is constitutively expressed in all examined tissues. In addition, our results indicate that MCs exposure significantly promotes the transcription of Hm-p38a in fish liver or kidney, suggesting that mitogen-activated protein kinase should also be the signal pathway of MCs hepatotoxicity in fish.

Keywords: Clone; Microcystins; Silver carp; Transcription; p38 MAPK.

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Bacterial Toxins / isolation & purification
  • Bacterial Toxins / toxicity*
  • Base Sequence
  • Carps / genetics*
  • Carps / metabolism
  • Cloning, Molecular
  • Conserved Sequence
  • Fish Proteins / agonists
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Gene Expression Regulation
  • Kidney / drug effects*
  • Kidney / enzymology
  • Liver / drug effects*
  • Liver / enzymology
  • Microcystins / isolation & purification
  • Microcystins / toxicity*
  • Microcystis / chemistry
  • Microcystis / pathogenicity
  • Open Reading Frames
  • Phylogeny
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Signal Transduction
  • Transcription, Genetic
  • p38 Mitogen-Activated Protein Kinases / genetics*
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Bacterial Toxins
  • Fish Proteins
  • Microcystins
  • microcystin
  • p38 Mitogen-Activated Protein Kinases