SUG1, a component of the 26 S proteasome, is an ATPase stimulated by specific RNAs.
SUG1 is an integral component of the 26 S proteasome. Belonging to a novel putative ATPase family, it shares four conserved motifs characteristic of ATP-dependent DNA/RNA helicases. Recombinant rat SUG1 (rSUG1) produced in Escherichia coli was highly purified and characterized in terms of its biochemical properties. The rSUG1 exhibited a Mg2+-dependent ATPase activity. The Km for ATP and Vmax of rSUG1 were 35 microM and 7 pmol of ATP/min/microg of protein, respectively. Both ATPase activity to release [32P]monophosphate and [32P]ATP-labeling activity were coordinately affected by cold ATP severely, GTP and UTP moderately, and CTP little. Interestingly, the rSUG1 ATPase activity was stimulated by poly(U) and poly(C), but not by poly(A), poly(G), or by any forms of DNAs tested. A UV cross-linking assay also indicated poly(U)- and poly(C)-stimulated labeling of rSUG1 with [alpha-32P]ATP. Moreover, the ATPase activity was facilitated by cellular poly(A)+ RNA, but not by poly(A)- RNA. RNA transcribed in vitro from cDNA encoding a b-Zip protein could stimulate the ATPase activity. This is the first report to demonstrate a specific RNA requirement for ATPase with respect to the proteasomal ATPases. Our present work suggests that SUG1 can specifically interact with protein-coding RNA (mRNA) and play some roles in mRNA metabolism.
Makino
Y
Y
Department of Biology, Faculty of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba-263, Japan.
Yamano
K
K
Kanemaki
M
M
Morikawa
K
K
Kishimoto
T
T
Shimbara
N
N
Tanaka
K
K
Tamura
T
T
D000251:Adenosine Triphosphatases / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D003546:Cysteine Endopeptidases / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006868:Hydrolysis; D009097:Multienzyme Complexes / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011066:Poly C / Q000494:pharmacology; D011072:Poly U / Q000494:pharmacology; D011131:Polyribonucleotides / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex; D012313:RNA / Q000494:pharmacology*; D012333:RNA, Messenger / Q000494:pharmacology; D051381:Rats; D011994:Recombinant Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism
D003432:Cross-Linking Reagents; D009097:Multienzyme Complexes; D011131:Polyribonucleotides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011072:Poly U; D011066:Poly C; D012313:RNA; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases