Structural and functional insights into Dom34, a key component of no-go mRNA decay.
The yeast protein Dom34 is a key component of no-go decay, by which mRNAs with translational stalls are endonucleolytically cleaved and subsequently degraded. However, the identity of the endoribonuclease is unknown. Homologs of Dom34, called Pelota, are broadly conserved in eukaryotes and archaea. To gain insights into the structure and function of Dom34/Pelota, we have determined the structure of Pelota from Thermoplasma acidophilum (Ta Pelota) and investigated the ribonuclease activity of Dom34/Pelota. The structure of Ta Pelota is tripartite, and its domain 1 has the RNA-binding Sm fold. We have discovered that Ta Pelota has a ribonuclease activity and that its domain 1 is sufficient for the catalytic activity. We also demonstrate that domain 1 of Dom34 has an endoribonuclease activity against defined RNA substrates containing a stem loop, which supports a direct catalytic role of yeast Dom34 in no-go mRNA decay.
Lee
Hyung Ho
HH
Department of Chemistry, College of Natural Sciences, Seoul National University, Seoul 151-747, Korea.
Kim
Youn-Sung
YS
Kim
Kyoung Hoon
KH
Heo
Inha
I
Kim
Sang Kyu
SK
Kim
Olesya
O
Kim
Hye Kyung
HK
Yoon
Ji Young
JY
Kim
Hyoun Sook
HS
Kim
Do Jin
DJ
Lee
Sang Jae
SJ
Yoon
Hye Jin
HJ
Kim
Soon Jong
SJ
Lee
Byung Gil
BG
Song
Hyun Kyu
HK
Kim
V Narry
VN
Park
Chung-Mo
CM
Suh
Se Won
SW
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D004722:Endoribonucleases; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010454:Peptide Termination Factors / Q000737:chemistry; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D012996:Solutions; D040681:Structural Homology, Protein; D013379:Substrate Specificity; D013822:Thermoplasma / Q000201:enzymology*
D019843:Archaeal Proteins; D018797:Cell Cycle Proteins; C489002:ETF1 protein, human; D010454:Peptide Termination Factors; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D012996:Solutions; C112808:Dom34 protein, S cerevisiae; D004722:Endoribonucleases; D012260:Ribonucleases