Alternative mRNA splicing of the novel GTPase Rab28 generates isoforms with different C-termini

Eur J Biochem. 1996 May 1;237(3):833-40. doi: 10.1111/j.1432-1033.1996.0833p.x.

Abstract

A novel ras-related gene (rab28) was identified by a PCR-based cloning approach and subsequent screening of rat fat cell and brain cDNA libraries. The deduced amino acid sequence of the cDNA is distantly related with members of the Rab family (31-33% sequence identity, mainly restricted to the six GTP-binding motifs). Cloning of the human homologue of Rab28 by a PCR-based approach revealed the existence of two isoforms (hRab28S, hRab28L) which differ only by a 95-bp insertion within the coding region. This insertion generates an alternative sequence of the 30 C-terminal amino acids of the protein. Both C-termini of the human homologues comprise farnesylation motifs, but differ strikingly in a stretch of 13 amino acids. By PCR, mRNA of hRab28S was detected in most tissues investigated (cortex, liver, kidney, skeletal muscle, adipose tissue, testis and urothelium), whereas hRab28L was predominant in testis. Recombinant Rab28 proteins showed specific binding of radiolabeled guanosine 5'-O-[gamma-thio]triphosphate and rapidly hydrolysed [alpha-32P]GTP; there was no difference in the GTP binding characteristics of the two isoforms hRab28S and hRab28L. It is suggested that the isoforms are derived from the same gene by alternative mRNA splicing, and that their functions differ in a parameter unrelated to its basic role as a GTPase.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / enzymology
  • Alternative Splicing
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Brain / enzymology
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • GTP Phosphohydrolases / chemistry
  • GTP Phosphohydrolases / genetics*
  • GTP Phosphohydrolases / metabolism*
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / genetics*
  • GTP-Binding Proteins / metabolism*
  • Humans
  • Isoenzymes / chemistry
  • Isoenzymes / genetics*
  • Isoenzymes / metabolism*
  • Male
  • Molecular Sequence Data
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism*
  • Rats
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Tissue Distribution
  • rab GTP-Binding Proteins*

Substances

  • DNA, Complementary
  • Isoenzymes
  • RNA, Messenger
  • Recombinant Proteins
  • GTP Phosphohydrolases
  • GTP-Binding Proteins
  • RAB28 protein, human
  • Rab28 protein, rat
  • rab GTP-Binding Proteins

Associated data

  • GENBANK/X78606
  • GENBANK/X94703