PMID- 26578560
OWN - NLM
STAT- MEDLINE
DCOM- 20160621
LR  - 20181113
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 43
IP  - 22
DP  - 2015 Dec 15
TI  - Structural basis for RNA-genome recognition during bacteriophage Qbeta replication.
PG  - 10893-906
LID - 10.1093/nar/gkv1212 [doi]
AB  - Upon infection of Escherichia coli by bacteriophage Qbeta, the virus-encoded 
      beta-subunit recruits host translation elongation factors EF-Tu and EF-Ts and 
      ribosomal protein S1 to form the Qbeta replicase holoenzyme complex, which is 
      responsible for amplifying the Qbeta (+)-RNA genome. Here, we use X-ray 
      crystallography, NMR spectroscopy, as well as sequence conservation, surface 
      electrostatic potential and mutational analyses to decipher the roles of the 
      beta-subunit and the first two oligonucleotide-oligosaccharide-binding domains of S1 
      (OB1-2) in the recognition of Qbeta (+)-RNA by the Qbeta replicase complex. We show how 
      three basic residues of the beta subunit form a patch located adjacent to the OB2 
      domain, and use NMR spectroscopy to demonstrate for the first time that OB2 is 
      able to interact with RNA. Neutralization of the basic residues by mutagenesis 
      results in a loss of both the phage infectivity in vivo and the ability of Qbeta 
      replicase to amplify the genomic RNA in vitro. In contrast, replication of 
      smaller replicable RNAs is not affected. Taken together, our data suggest that 
      the beta-subunit and protein S1 cooperatively bind the (+)-stranded Qbeta genome during 
      replication initiation and provide a foundation for understanding template 
      discrimination during replication initiation.
CI  - (c) The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic 
      Acids Research.
FAU - Gytz, Heidi
AU  - Gytz H
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark.
FAU - Mohr, Durita
AU  - Mohr D
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark.
FAU - Seweryn, Paulina
AU  - Seweryn P
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark.
FAU - Yoshimura, Yuichi
AU  - Yoshimura Y
AD  - Interdisciplinary Nanoscience Centre (iNANO) and Department of Chemistry, Aarhus 
      University, DK-8000 Aarhus C, Denmark.
FAU - Kutlubaeva, Zarina
AU  - Kutlubaeva Z
AD  - Institute of Protein Research of the Russian Academy of Sciences, Pushchino, 
      Moscow Region 142290, Russia.
FAU - Dolman, Fleur
AU  - Dolman F
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark.
FAU - Chelchessa, Bosene
AU  - Chelchessa B
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark.
FAU - Chetverin, Alexander B
AU  - Chetverin AB
AD  - Institute of Protein Research of the Russian Academy of Sciences, Pushchino, 
      Moscow Region 142290, Russia.
FAU - Mulder, Frans A A
AU  - Mulder FA
AD  - Interdisciplinary Nanoscience Centre (iNANO) and Department of Chemistry, Aarhus 
      University, DK-8000 Aarhus C, Denmark.
FAU - Brodersen, Ditlev E
AU  - Brodersen DE
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark.
FAU - Knudsen, Charlotte R
AU  - Knudsen CR
AD  - Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus 
      C, Denmark crk@mbg.au.dk.
LA  - eng
SI  - PDB/4R71
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151117
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Protein Subunits)
RN  - 0 (RNA, Viral)
RN  - 0 (Ribosomal Proteins)
RN  - 0 (ribosomal protein S1, E coli)
RN  - EC 2.7.7.- (Q beta Replicase)
SB  - IM
MH  - Allolevivirus/genetics/*physiology
MH  - Dimerization
MH  - Escherichia coli Proteins/chemistry/metabolism
MH  - *Genome, Viral
MH  - Mutation
MH  - Protein Binding
MH  - Protein Structure, Tertiary
MH  - Protein Subunits/chemistry/genetics/metabolism
MH  - Q beta Replicase/*chemistry/genetics/metabolism
MH  - RNA, Viral/biosynthesis/metabolism
MH  - Ribosomal Proteins/chemistry/metabolism
MH  - *Virus Replication
PMC - PMC4678825
EDAT- 2015/11/19 06:00
MHDA- 2016/06/22 06:00
PMCR- 2015/11/17
CRDT- 2015/11/19 06:00
PHST- 2015/10/28 00:00 [accepted]
PHST- 2015/09/10 00:00 [received]
PHST- 2015/11/19 06:00 [entrez]
PHST- 2015/11/19 06:00 [pubmed]
PHST- 2016/06/22 06:00 [medline]
PHST- 2015/11/17 00:00 [pmc-release]
AID - gkv1212 [pii]
AID - 10.1093/nar/gkv1212 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2015 Dec 15;43(22):10893-906. doi: 10.1093/nar/gkv1212. Epub 
      2015 Nov 17.