Molecular characterization and function of a PTEN gene from Litopenaeus vannamei after Vibrio alginolyticus challenge

Dev Comp Immunol. 2016 Jun:59:77-88. doi: 10.1016/j.dci.2016.01.004. Epub 2016 Feb 4.

Abstract

PTEN, a tumor suppressor gene, suppresses cell survival, growth, apoptosis, cell migration and DNA damage repair by inhibiting the PI3K/AKT signaling pathway. In this study, the full-length Litopenaeus vannamei PTEN (LvPTEN) cDNA was obtained, containing a 5'UTR of 59bp, an ORF of 1269bp and a 3'UTR of 146bp besides the poly (A) tail. The PTEN gene encoded a protein of 422 amino acids with an estimated molecular mass of 48.3 KDa and a predicted isoelectric point (pI) of 7.6. Subcellular localization analysis revealed that LvPTEN was distributed in both cytoplasm and nucleus, and the tissue distribution patterns showed that LvPTEN was ubiquitously expressed in all the examined tissues. Vibrio alginolyticus challenge induced upregulation of LvPTEN expression. Moreover, RNAi knock-down of LvPTEN in vivo significantly increased the expression of LvAKT mRNA, while reducing that of the downstream apoptosis genes LvP53 and LvCaspase3. LvPTEN knock-down also caused a sharp increase in cumulative mortality, bacterial numbers, and DNA damage in the hemolymph of L. vannamei following V. alginolyticus challenge, together with a sharp decrease in the total hemocyte count (THC). These results suggested that LvPTEN may participate in apoptosis via the PI3K/AKT signaling pathway in L. vannamei, and play an important role in shrimp innate immunity.

Keywords: Apoptosis; Litopenaeus vannamei; LvPTEN; RNAi; Shrimp immunity; Vibrio alginolyticus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis / immunology
  • Base Sequence
  • Caspase 3 / biosynthesis
  • Cloning, Molecular
  • DNA Damage / genetics
  • DNA, Complementary / genetics
  • Hemocytes / immunology
  • Hemolymph / cytology
  • Hemolymph / microbiology
  • Immunity, Innate / immunology*
  • PTEN Phosphohydrolase / genetics*
  • PTEN Phosphohydrolase / immunology
  • Penaeidae / genetics
  • Penaeidae / immunology*
  • Penaeidae / microbiology
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphoinositide-3 Kinase Inhibitors
  • Proto-Oncogene Proteins c-akt / biosynthesis
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Sequence Alignment
  • Tumor Suppressor Protein p53 / biosynthesis
  • Vibrio Infections / immunology*
  • Vibrio Infections / microbiology
  • Vibrio alginolyticus / immunology*

Substances

  • DNA, Complementary
  • Phosphoinositide-3 Kinase Inhibitors
  • RNA, Small Interfering
  • Tumor Suppressor Protein p53
  • Proto-Oncogene Proteins c-akt
  • PTEN Phosphohydrolase
  • Caspase 3