TY - JOUR
T1 - YxiN is a modular protein combining a DExD/H core and a specific RNA-binding domain
AU - Karginov, Fedor V.
AU - Caruthers, Jonathan M.
AU - Hu, Yao Xiong
AU - McKay, David B.
AU - Uhlenbeck, Olke C.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2005/10/21
Y1 - 2005/10/21
N2 - DExD/H proteins, typically described as RNA helicases, participate in rearrangement of RNA-RNA and possibly RNA-protein complexes in the cell. Aside from the conserved DExD/H core, members of this protein family often contain N- and C-terminal extensions that are responsible for additional functions. The Bacillus subtilis DEx D/H-box protein YxiN and its Escherichia coli ortholog DbpA contain an ∼80 amino acid C-terminal extension that has been proposed to specifically interact with a region of 23 S ribosomal RNA including hairpin 92. In this study, the DExD/H-box core and the C-terminal domain of YxiN were expressed and characterized as separate proteins. The isolated DExD/H-box core, YxCat, had weak, nonspecific RNA binding activity and showed RNA-stimulated ATPase activity with a K m(ATP) that resembled several non-specific DExD/ H proteins. The isolated C-terminal domain, YxRBD, bound RNA with the high affinity and specificity seen with full-length YxiN. Thus, YxiN is a modular protein combining the activities of the YxCat and YxRBD domains. Footprinting of YxiN and YxRBD on a 172-nucleotide fragment of 23 S rRNA was used to identify the sites of interaction of the C-terminal and helicase domains with the RNA.
AB - DExD/H proteins, typically described as RNA helicases, participate in rearrangement of RNA-RNA and possibly RNA-protein complexes in the cell. Aside from the conserved DExD/H core, members of this protein family often contain N- and C-terminal extensions that are responsible for additional functions. The Bacillus subtilis DEx D/H-box protein YxiN and its Escherichia coli ortholog DbpA contain an ∼80 amino acid C-terminal extension that has been proposed to specifically interact with a region of 23 S ribosomal RNA including hairpin 92. In this study, the DExD/H-box core and the C-terminal domain of YxiN were expressed and characterized as separate proteins. The isolated DExD/H-box core, YxCat, had weak, nonspecific RNA binding activity and showed RNA-stimulated ATPase activity with a K m(ATP) that resembled several non-specific DExD/ H proteins. The isolated C-terminal domain, YxRBD, bound RNA with the high affinity and specificity seen with full-length YxiN. Thus, YxiN is a modular protein combining the activities of the YxCat and YxRBD domains. Footprinting of YxiN and YxRBD on a 172-nucleotide fragment of 23 S rRNA was used to identify the sites of interaction of the C-terminal and helicase domains with the RNA.
UR - http://www.scopus.com/inward/record.url?scp=27444442034&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=27444442034&partnerID=8YFLogxK
U2 - 10.1074/jbc.M506815200
DO - 10.1074/jbc.M506815200
M3 - Article
C2 - 16118224
AN - SCOPUS:27444442034
VL - 280
SP - 35499
EP - 35505
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 42
ER -