PMID- 19710136
OWN - NLM
STAT- MEDLINE
DCOM- 20091123
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 83
IP  - 21
DP  - 2009 Nov
TI  - Ribose 2'-O methylation of the vesicular stomatitis virus mRNA cap precedes and 
      facilitates subsequent guanine-N-7 methylation by the large polymerase protein.
PG  - 11043-50
LID - 10.1128/JVI.01426-09 [doi]
AB  - During conventional mRNA cap formation, two separate methyltransferases 
      sequentially modify the cap structure, first at the guanine-N-7 (G-N-7) position 
      and subsequently at the ribose 2'-O position. For vesicular stomatitis virus 
      (VSV), a prototype of the nonsegmented negative-strand RNA viruses, the two 
      methylase activities share a binding site for the methyl donor 
      S-adenosyl-l-methionine and are inhibited by individual amino acid substitutions 
      within the C-terminal domain of the large (L) polymerase protein. This led to the 
      suggestion that a single methylase domain functions for both 2'-O and G-N-7 
      methylations. Here we report a trans-methylation assay that recapitulates both 
      ribose 2'-O and G-N-7 modifications by using purified recombinant L and in 
      vitro-synthesized RNA. Using this assay, we demonstrate that VSV L typically 
      modifies the 2'-O position of the cap prior to the G-N-7 position and that G-N-7 
      methylation is diminished by pre-2'-O methylation of the substrate RNA. Amino 
      acid substitutions in the C terminus of L that prevent all cap methylation in 
      recombinant VSV (rVSV) partially retain the ability to G-N-7 methylate a 
      pre-2'-O-methylated RNA, therefore uncoupling the effect of substitutions in the 
      C terminus of the L protein on the two methylations. In addition, we show that 
      the 2'-O and G-N-7 methylase activities act specifically on RNA substrates that 
      contain the conserved elements of a VSV mRNA start at the 5' terminus. This study 
      provides new mechanistic insights into the mRNA cap methylase activities of VSV 
      L, demonstrates that 2'-O methylation precedes and facilitates subsequent G-N-7 
      methylation, and reveals an RNA sequence and length requirement for the two 
      methylase activities. We propose a model of regulation of the activity of the C 
      terminus of L protein in 2'-O and G-N-7 methylation of the cap structure.
FAU - Rahmeh, Amal A
AU  - Rahmeh AA
AD  - Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 
      Longwood Ave., Boston, MA 02115, USA.
FAU - Li, Jianrong
AU  - Li J
FAU - Kranzusch, Philip J
AU  - Kranzusch PJ
FAU - Whelan, Sean P J
AU  - Whelan SP
LA  - eng
GR  - R01 AI059371-05/AI/NIAID NIH HHS/United States
GR  - R37 AI059371/AI/NIAID NIH HHS/United States
GR  - AI059371/AI/NIAID NIH HHS/United States
GR  - R01 AI059371/AI/NIAID NIH HHS/United States
GR  - U54 AI057159/AI/NIAID NIH HHS/United States
GR  - T32 AI007245/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20090826
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (RNA Caps)
RN  - 0 (RNA, Messenger)
RN  - 0 (Viral Proteins)
RN  - 5Z93L87A1R (Guanine)
RN  - 681HV46001 (Ribose)
RN  - EC 2.1.1.- (Methyltransferases)
RN  - EC 2.7.7.6 (DNA-Directed RNA Polymerases)
RN  - EC 2.7.7.6 (L polymerase protein, Vesicular stomatitis-Indiana virus)
SB  - IM
MH  - Amino Acid Substitution
MH  - Base Sequence
MH  - Binding Sites
MH  - DNA-Directed RNA Polymerases/antagonists & inhibitors/genetics/*metabolism
MH  - Guanine/*metabolism
MH  - Methylation
MH  - Methyltransferases/antagonists & inhibitors/metabolism
MH  - RNA Caps/genetics/*metabolism
MH  - RNA, Messenger/genetics/*metabolism
MH  - Ribose/*metabolism
MH  - Vesiculovirus/*genetics/*metabolism
MH  - Viral Proteins/antagonists & inhibitors/genetics/*metabolism
PMC - PMC2772757
EDAT- 2009/08/28 09:00
MHDA- 2009/12/16 06:00
PMCR- 2010/05/01
CRDT- 2009/08/28 09:00
PHST- 2009/08/28 09:00 [entrez]
PHST- 2009/08/28 09:00 [pubmed]
PHST- 2009/12/16 06:00 [medline]
PHST- 2010/05/01 00:00 [pmc-release]
AID - JVI.01426-09 [pii]
AID - 1426-09 [pii]
AID - 10.1128/JVI.01426-09 [doi]
PST - ppublish
SO  - J Virol. 2009 Nov;83(21):11043-50. doi: 10.1128/JVI.01426-09. Epub 2009 Aug 26.