Structure, organization and expression of common carp (Cyprinus carpio L.) SLP-76 gene

Fish Shellfish Immunol. 2008 May;24(5):530-41. doi: 10.1016/j.fsi.2007.10.011. Epub 2007 Nov 5.

Abstract

SLP-76 is an important member of the SLP-76 family of adapters, and it plays a key role in TCR signaling and T cell function. Partial cDNA sequence of SLP-76 of common carp (Cyprinus carpio L.) was isolated from thymus cDNA library by the method of suppression subtractive hybridization (SSH). Subsequently, the full length cDNA of carp SLP-76 was obtained by means of 3' RACE and 5' RACE, respectively. The full length cDNA of carp SLP-76 was 2007 bp, consisting of a 5'-terminal untranslated region (UTR) of 285 bp, a 3'-terminal UTR of 240 bp, and an open reading frame of 1482 bp. Sequence comparison showed that the deduced amino acid sequence of carp SLP-76 had an overall similarity of 34-73% to that of other species homologues, and it was composed of an NH2-terminal domain, a central proline-rich domain, and a C-terminal SH2 domain. Amino acid sequence analysis indicated the existence of a Gads binding site R-X-X-K, a 10-aa-long sequence which binds to the SH3 domain of LCK in vitro, and three conserved tyrosine-containing sequence in the NH2-terminal domain. Then we used PCR to obtain a genomic DNA which covers the entire coding region of carp SLP-76. In the 9.2k-long genomic sequence, twenty one exons and twenty introns were identified. RT-PCR results showed that carp SLP-76 was expressed predominantly in hematopoietic tissues, and was upregulated in thymus tissue of four-month carp compared to one-year old carp. RT-PCR and virtual northern hybridization results showed that carp SLP-76 was also upregulated in thymus tissue of GH transgenic carp at the age of four-months. These results suggest that the expression level of SLP-76 gene may be related to thymocyte development in teleosts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / chemistry
  • Adaptor Proteins, Signal Transducing / genetics*
  • Aging / genetics
  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Carps / genetics*
  • Cloning, Molecular
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Gene Expression Profiling
  • Gene Expression Regulation / genetics*
  • Humans
  • Molecular Sequence Data
  • Phosphoproteins / chemistry
  • Phosphoproteins / genetics*
  • Phylogeny
  • Sequence Alignment
  • Thymus Gland / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • Fish Proteins
  • Phosphoproteins
  • SLP-76 signal Transducing adaptor proteins

Associated data

  • GENBANK/EU090723
  • GENBANK/EU090724