ACreTranscription Fidelity Reporter Identifies GreA as a Major RNA Proofreading Factor inEscherichia coli.
We made a coupled genetic reporter that detects rare transcription misincorporation errors to measure RNA polymerase transcription fidelity inEscherichia coliUsing this reporter, we demonstratedin vivothat the transcript cleavage factor GreA, but not GreB, is essential for proofreading of a transcription error where a riboA has been misincorporated instead of a riboG. AgreAmutant strain had more than a 100-fold increase in transcription errors relative to wild-type or agreBmutant. However, overexpression of GreB in ΔgreAcells reduced the misincorporation errors to wild-type levels, demonstrating that GreB at high concentration could substitute for GreA in RNA proofreading activityin vivo.
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Mikhail G
MG
Bubunenko
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Carolyn B
CB
Court
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Alison J
AJ
Rattray
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Deanna R
DR
Gotte
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Maria L
ML
Kireeva
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Jorge A
JA
Irizarry-Caro
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702.
Xintian
X
Li
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702 jind@mail.nih.gov courtd@mail.nih.gov strathej@mail.nih.gov kashlevm@mail.nih.gov.
Ding J
DJ
Jin
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702 jind@mail.nih.gov courtd@mail.nih.gov strathej@mail.nih.gov kashlevm@mail.nih.gov.
Donald L
DL
Court
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702 jind@mail.nih.gov courtd@mail.nih.gov strathej@mail.nih.gov kashlevm@mail.nih.gov.
Jeffrey N
JN
Strathern
The RNA Biology Laboratory, National Cancer Institute-Frederick, Center for Cancer Research, Maryland 21702 jind@mail.nih.gov courtd@mail.nih.gov strathej@mail.nih.gov kashlevm@mail.nih.gov.
Mikhail
M
Kashlev
D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter / Q000235:genetics*; D010445:Peptide Elongation Factors; D011401:Promoter Regions, Genetic; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D035602:Transcriptional Elongation Factors / Q000235:genetics*
D029968:Escherichia coli Proteins; C079668:GreA protein, E coli; C079667:GreB protein, E coli; D010445:Peptide Elongation Factors; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; D012313:RNA; D012321:DNA-Directed RNA Polymerases
Cre/lox fidelity reporter; Gre proteins; RNA polymerase; transcription errors