cDNA cloning and characterization of a human proteasomal modulator subunit, p27 (PSMD9).
We have employed cDNA cloning to deduce the complete primary structure of a new subunit, designated p27, of the modulator trimer complex that stimulates the association of the PA700 regulator with the catalytic 20S proteasome to form the ATP-dependent active 26S proteasome. We found two distinct cDNAs encoding two highly homologous proteins except in the C-terminal region, which are termed tentatively p27-1 and p27-2. The short p27-2 cDNA has a deletion of 65 bp near the 3'-end region of the long p27-1 cDNA, which encodes a large protein with an extended C-terminal region, designated p27-L, whereas the long p27-1 cDNA encodes a small protein named p27-S. The polypeptides of p27-L and p27-S consist of 223 and 209 amino acid residues with calculated molecular masses of 24,852 and 22,764 and isoelectric points of 6.50 and 5.28, respectively. Immunoblot analysis with anti-p27 antibody revealed that p27, together with two other ATPase components, TBP1 and p42, was associated with not only the modulator complex but also significantly with the 26S proteasome complex, suggesting that the three are common/sharing subunits in these two complexes. By the fluorescence in situ hybridization method, the p27 (PSMD9) gene was mapped to the q24.2-q24.3 band of human chromosome 12. Computer-assisted homology analysis revealed the high sequence similarities of p27-L with a possible counterpart in Caenorhabditis elegans and Saccharomyces cerevisiae whose function is yet unknown, the yeast gene that is here termed NAS2 (non-ATPase subunit 2). Disruption of NAS2 had no effect on cell viability, indicating that the subunit is not essential for proliferation of yeast cells.
Watanabe
T K
TK
Otsuka GEN Research Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan.
Saito
A
A
Suzuki
M
M
Fujiwara
T
T
Takahashi
E
E
Slaughter
C A
CA
DeMartino
G N
GN
Hendil
K B
KB
Chung
C H
CH
Tanahashi
N
N
Tanaka
K
K
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D017173:Caenorhabditis elegans / Q000235:genetics; D002874:Chromosome Mapping; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases; D018076:DNA, Complementary; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D046988:Proteasome Endopeptidase Complex; D011506:Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D005656:Fungal Proteins; D009097:Multienzyme Complexes; C114475:NAS2 protein, S cerevisiae; C085834:PA700 proteasome activator; C114476:PSMD9 protein, human; D011506:Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases