Molecular cloning of two types of cDNA encoding subunit RC6-I of rat proteasomes

Runzhou Ni, Yumiko Tomita, Fuminori Tokunaga, T. Jake Liang, Chiseko Noda, Akira Ichihara, Keiji Tanaka

Research output: Contribution to journalArticle

Abstract

A new subunit, named RC6-I, of the rat 20 S proteasome was purified and the partial amino acid sequences of several peptide fragments obtained by digestion with lysyl-endopeptidase were determined by Edman degradation. Amplification of cDNAs encoding RC6-I by the polymerase chain reaction (PCR) technique revealed two types of cDNA, tentatively designated as RC6-IL and RC6-IS in order of size. The nucleotide sequences of the two cDNAs are identical except that RC6-IL contains an insertion of 18 nucleotides in the coding region compared with RC6-IS. The polypeptide predicted from the open reading frame of RC6-IS cDNA consists of 248 amino acid residues with a calculated molecular weight of 27783. These values are consistent with those obtained by protein chemical analyses. Computer-assisted homology analysis showed that RC6-I belongs to the α-type subfamily of the proteasome gene family, which shows similarity to the α-subunit of the archaebacterium Thermoplasma acidophilum proteasome, and that the 18 nucleotide insert, encoding six amino acid residues, VVASVS, appears to be unique to RC6-IL, because this motif has not been conserved in any other α-type subunit. By reverse transcription (RT)-PCR analysis, the mRNAs for both RC6-IL and RC6-IS were found in all the rat tissues examined. These results suggest that proteasomes are present as a heterogeneous population, possibly for acquisition of diversity of functions.

Original languageEnglish (US)
Pages (from-to)45-52
Number of pages8
JournalBBA - Gene Structure and Expression
Volume1264
Issue number1
DOIs
StatePublished - Oct 17 1995
Externally publishedYes

Fingerprint

Cloning
Molecular Cloning
Proteasome Endopeptidase Complex
Rats
Complementary DNA
Nucleotides
lysyl endopeptidase
Polymerase chain reaction
Amino Acids
Polymerase Chain Reaction
Peptide Fragments
Archaea
Transcription
Open Reading Frames
Reverse Transcription
Amplification
Digestion
Amino Acid Sequence
Genes
Molecular Weight

Keywords

  • cDNA cloning
  • Multicatalytic proteinase complex
  • Proteasome
  • Subunit RC6-I

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics
  • Molecular Biology
  • Structural Biology

Cite this

Ni, R., Tomita, Y., Tokunaga, F., Jake Liang, T., Noda, C., Ichihara, A., & Tanaka, K. (1995). Molecular cloning of two types of cDNA encoding subunit RC6-I of rat proteasomes. BBA - Gene Structure and Expression, 1264(1), 45-52. https://doi.org/10.1016/0167-4781(95)00113-U

Molecular cloning of two types of cDNA encoding subunit RC6-I of rat proteasomes. / Ni, Runzhou; Tomita, Yumiko; Tokunaga, Fuminori; Jake Liang, T.; Noda, Chiseko; Ichihara, Akira; Tanaka, Keiji.

In: BBA - Gene Structure and Expression, Vol. 1264, No. 1, 17.10.1995, p. 45-52.

Research output: Contribution to journalArticle

Ni, R, Tomita, Y, Tokunaga, F, Jake Liang, T, Noda, C, Ichihara, A & Tanaka, K 1995, 'Molecular cloning of two types of cDNA encoding subunit RC6-I of rat proteasomes', BBA - Gene Structure and Expression, vol. 1264, no. 1, pp. 45-52. https://doi.org/10.1016/0167-4781(95)00113-U
Ni, Runzhou ; Tomita, Yumiko ; Tokunaga, Fuminori ; Jake Liang, T. ; Noda, Chiseko ; Ichihara, Akira ; Tanaka, Keiji. / Molecular cloning of two types of cDNA encoding subunit RC6-I of rat proteasomes. In: BBA - Gene Structure and Expression. 1995 ; Vol. 1264, No. 1. pp. 45-52.
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