Identification, characterization and expression analysis of tumor suppressor protein p53 from pufferfish (Takifugu obscurus) after the Vibrio alginolyticus challenge

Fish Shellfish Immunol. 2016 Dec:59:312-322. doi: 10.1016/j.fsi.2016.10.040. Epub 2016 Nov 1.

Abstract

The tumor suppressor protein p53 plays a critical role in cell cycle, apoptosis and DNA repair. In this study, the full-length pufferfish p53 (Pf-p53) was obtained, containing an open reading frame of 1095 bp, a 5'UTR of 157 bp and a 3'UTR of 285 bp with a poly (A) tail. The Pf-p53 encoded a polypeptide of 364 amino acids with a theoretical isoelectric point of 8.03 and predicted molecular weight of 40.6 kDa. Pf-p53 was ubiquitously expressed in various tissues with a high-level expression in kidney, liver and gill. Vibrio alginolyticus challenge could induce ROS production and disrupt Ca2+ homeostasis, subsequently leading to the induction of DNA damage and apoptosis, while the Vibrio alginolyticus-induced oxidative stress can also increase the non-specific immunity. The pufferfish challenged with Vibrio alginolyticus showed a sharp increase of Pf-p53 transcript in liver. Subcellular localization analysis revealed that Pf-p53 was primarily localized in nucleus. Furthermore, overexpression of Pf-p53 in Hela cells could inhibit cell proliferation and the transcriptional activities of the NF-ĸB promoter. Taken together, our results indicated that Pf-p53 may play an important role in the immune response to Vibrio alginolyticus challenge.

Keywords: Apoptosis; Gene expression; Takifugu obscurus; Vibrio alginolyticus; p53.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Fish Diseases / genetics
  • Fish Diseases / immunology*
  • Fish Diseases / microbiology
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Gene Expression Regulation / immunology*
  • Immunity, Innate / genetics*
  • Phylogeny
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Random Allocation
  • Sequence Alignment / veterinary
  • Takifugu*
  • Tissue Distribution
  • Tumor Suppressor Protein p53 / chemistry
  • Tumor Suppressor Protein p53 / genetics*
  • Tumor Suppressor Protein p53 / metabolism
  • Up-Regulation
  • Vibrio Infections / genetics
  • Vibrio Infections / immunology
  • Vibrio Infections / microbiology
  • Vibrio Infections / veterinary*
  • Vibrio alginolyticus / physiology

Substances

  • DNA, Complementary
  • Fish Proteins
  • RNA, Messenger
  • Tumor Suppressor Protein p53