cDNA cloning and characterization of the protein encoded by RD, a gene located in the class III region of the human major histocompatibility complex.
The RD gene, initially defined in the mouse, has been mapped between the Bf and C4A genes in the human major histocompatibility complex class III region. Using the mouse cDNA as a probe, we isolated and sequenced human RD cDNA clones. The composite nucleotide sequence consisted of 1301 nucleotides, excluding a poly(A) tail at the 3' end. It contained a single open reading frame encoding a polypeptide of 380 amino acid residues with a calculated molecular mass of 42274 Da. The most striking structural feature of the deduced amino acid sequence is a region consisting entirely of 24 tandem repeats of an Arg-Asp (or Glu) dipeptide. The human RD cDNA was expressed in Escherichia coli as a fusion protein with glutathione S-transferase and used to produce antisera in rabbits. Western blot analysis and immunoprecipitation of lysates of biosynthetically labelled HeLa cells indicated that RD is a 44 kDa nuclear protein.
Cheng
J
J
Department of Medicine, University of Alabama, Birmingham 35294.
Macon
K J
KJ
Volanakis
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007163:Immunosorbent Techniques; D008285:Major Histocompatibility Complex / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors
D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C498268:Rdbp protein, mouse; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C118437:negative elongation factor; D004247:DNA; D005982:Glutathione Transferase