cDNA cloning of rat proteasome subunit RC10‐II, assumed to be responsible for trypsin‐like catalytic activity

Abstract

The nucleotide sequence of a cDNA that encodes a new subunit, named RClO-II, of the 20s proteasome of rat embryonic brain has been determined. The polypeptide predicted from the open reading frame consists of 205 amino acid residues with a calculated molecular weight of 22,965 and isoelectric point of 6.15. Computer analysis showed that RClO-II belongs to the /I-type subgroup of proteasomes, differing clearly from a-type subunits of the proteasome gene family. The primary structure of RClO-II was found to contain the partial amino acid sequences of several fragments of subunit 13, which has a cysteinyl residue critical for the trypsin-like catalytic activity, as reported by L.R. Dick et al. [Biochemistry 31, 7347-7355, 19921, suggesting that RClO-II is a proteasomal subunit necessary for the expression of tryptic activity.

PRE5 and PRE6, the Last Missing Genes Encoding 20S Proteasome Subunits from Yeast? Indication for a Set of 14 Different Subunits in the Eukaryotic Proteasome Core

“…Cloning of PRE5 and PRE6. The 12 so far characterized genes encoding subunits of the 5. cerevisiae 20S proteasome can be divided into five a-type genes [PRS1, PRS2 (Fujiwara et al, 1990), PUP2 (Georgatsou et al, 1992), " Y7", and "Y13" (Emori et al, 1991)] and seven /3-type genes [PRE1 (Heinemeyer et al, 1991), PRE2 (Heinemeyer et al, 1993), PRE3 (Enenkel et al, 1994), PRE4 (Hilt et al, 1993), PRS3 (Lee etal., 1992), PUPl (Haffter & Fox, 1991), and the originally unidentified putative proteasomal gene (Basile et al, 1992) here referred to as PUP3, coding for a protein with striking homology to the recently characterized rat protesomal subunit RC10II (Nishimura et al, 1993b) and peptide sequences derived from the bovine proteasomal subunit 8 (Dick et al, 1992)]. When the primary structures of all known eukaryotic proteasomal proteins were compared with each other, each of the 12 yeast subunits fell into a separate of a total of 14 subgroups.…”

“…Development or tissue specific variations among proteasome species or even a diversity within one cell is not excluded by this rule. Subunit substitutions and modifications may be a widespread mechanism in higher eukaryotes to modulate the et al, 1990b), rn-C9 (Kumatori et al, 1990), rn-i, rn-f, and rn-LMP2 (Tamura et al, 1992), m-LMP7 (Aki et al, 1992), rn-RN3 (Thomson et al, 1993), rn-C71 (Nishimura et al, 1993a), m-ClOII (Nishimura et al, 1993b), sc-Y7 and sc-Y13 (Emori etal., 1991), sc-PRSl and sc-PRS2 (Fujiwara et al, 1990),sc-PUP2 (Georgatsou etal., 1992,sc-PRE3 (Enenkel et al, 1994), sc-PRE2 (Heinemeyer etal., 1993,sc-PUPl (Haffter & Fox, 1991),sc-PREl (Heinemeyer et al, 1991), sc-PRE4 (Hilt et al, 1993), sc-PUP3 (Basile et al, 1992,sc-PRS3 (Leeet al, 1992), sp-ptsl (Stone, Tanaka, Ichihara, Goebl, and Yanagida, unpublished; EMBL Data Base accession numberD13094),xl-C3 (Fujiietal., 1991),xl-C8 andxl-C9-l (Fujii etal., 1993, and xl-0 (Van Riel & Martens, 1991). The dendrogram was created by the CLUSTAL multiple sequence alignment program (Higgins & Sharp, 1988) using the DNASTAR "Lasergene" software.…”

The proteasomal system

“…HsC10-II 22.9 Nishimura et al (1993) and Nothwang et al (1994) b4 C11/Pre1 HsC7-I 22.8 Nothwang et al (1994) and Reidlinger et al (1997) Scheffler et al (2008), Egerer et al (2006), Caudill et al (2006) and Heink et al, 2006 b6 C5/Prs3 HsC5 26.5 (23.3) Trachtulec et al (1997) and Rodriguez-Vilarino et al (2000) b7…”

Identification of leptin-induced transcripts in the mouse hypothalamus.

“…Thus, it is possible that, although we see no significant induction of LIG 2 in the hypothalamus in vivo, LIG 2 induction by OB-R L in T-cells may have physiological significance. LIG 3 encodes a subunit of the 20s proteosome of the ␤-type subgroup of proteosomes and contains a critical cysteinyl residue necessary for the trypsin-like catalytic activity of this proteosome (39). Consequently, LIG 3 expression may function in neuropeptide processing.…”