Functional Analysis of BipA in E. coli Reveals the Natural Plasticity of 50S Subunit Assembly.
GTPase
TypA
YchF
protein synthesis
ribosome biogenesis
Journal
Journal of molecular biology
ISSN: 1089-8638
Titre abrégé: J Mol Biol
Pays: Netherlands
ID NLM: 2985088R
Informations de publication
Date de publication:
04 09 2020
04 09 2020
Historique:
received:
15
06
2020
revised:
20
07
2020
accepted:
20
07
2020
pubmed:
28
7
2020
medline:
10
3
2021
entrez:
26
7
2020
Statut:
ppublish
Résumé
BipA is a conserved translational GTPase of bacteria recently implicated in ribosome biogenesis. Here we show that Escherichia coli ΔbipA cells grown at suboptimal temperature accumulate immature large subunit particles missing several proteins. These include L17 and L17-dependent binders, suggesting that structural block 3 of the subunit folds late in the assembly process. Parallel analysis of the control strain revealed accumulation of nearly identical intermediates, albeit at lower levels, suggesting qualitatively similar routes of assembly. This came as a surprise, because earlier analogous studies of wild-type E. coli showed early binding of L17. Further investigation showed that the main path of 50S assembly differs depending on conditions of growth. Either supplementation of the media with lysine and arginine or suboptimal temperature appears to delay block 3 folding, demonstrating the flexible nature of the assembly process. We also show that the variant BipA-H78A fails to rescue phenotypes of the ΔbipA strain, indicating a critical role for GTP hydrolysis in BipA function. In fact, BipA-H78A confers a dominant negative phenotype in wild-type cells. Controlled production of BipA-H78A causes accumulation of 70S monosomes at the expense of polysomes, suggesting that the growth defect stems from a shutdown of translation.
Identifiants
pubmed: 32710983
pii: S0022-2836(20)30460-5
doi: 10.1016/j.jmb.2020.07.013
pmc: PMC7502552
mid: NIHMS1615225
pii:
doi:
Substances chimiques
Escherichia coli Proteins
0
Guanosine Triphosphate
86-01-1
GTP Phosphohydrolases
EC 3.6.1.-
typA protein, E coli
EC 3.6.1.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
5259-5272Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM072528
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM086252
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.
Références
J Bacteriol. 2008 Dec;190(23):7675-83
pubmed: 18820021
RNA. 2002 Jul;8(7):878-89
pubmed: 12166643
Science. 2010 Oct 29;330(6004):673-7
pubmed: 21030658
RNA. 2005 May;11(5):728-38
pubmed: 15840820
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4713-7
pubmed: 4612527
J Chromatogr A. 2002 Dec 6;979(1-2):233-9
pubmed: 12498253
J Mol Biol. 2003 Sep 19;332(3):689-99
pubmed: 12963376
Gene. 1991 Apr;100:195-9
pubmed: 2055470
Mol Microbiol. 1998 Apr;28(2):265-79
pubmed: 9622352
EcoSal Plus. 2004 Dec;1(1):
pubmed: 26443366
J Bacteriol. 2015 May;197(10):1819-27
pubmed: 25777676
Nature. 2008 Oct 30;455(7217):1268-72
pubmed: 18784650
J Bacteriol. 1995 Jul;177(14):4121-30
pubmed: 7608087
Nucleic Acids Res. 2013 Feb 1;41(4):2609-20
pubmed: 23293003
FASEB J. 1993 Jan;7(1):177-80
pubmed: 7916699
Nucleic Acids Res. 2014 Dec 1;42(21):13370-83
pubmed: 25378333
Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):999-1003
pubmed: 9448274
Cell. 2016 Dec 1;167(6):1610-1622.e15
pubmed: 27912064
Proc Natl Acad Sci U S A. 1982 Feb;79(3):729-33
pubmed: 7038683
J Biol Chem. 2018 Nov 23;293(47):18404-18419
pubmed: 30305394
Proc Natl Acad Sci U S A. 2017 Sep 26;114(39):E8165-E8173
pubmed: 28894000
Biochem J. 2016 Aug 1;473(15):2315-29
pubmed: 27470594
Mol Cell Proteomics. 2006 Dec;5(12):2252-62
pubmed: 16920818
Mol Syst Biol. 2006;2:2006.0008
pubmed: 16738554
Nucleic Acids Res. 2016 Feb 29;44(4):1952-61
pubmed: 26733579
Proc Natl Acad Sci U S A. 2016 Oct 11;113(41):E6126-E6134
pubmed: 27698115
Mol Microbiol. 2003 Apr;48(2):507-21
pubmed: 12675808
Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):7884-8
pubmed: 8058729
Electrophoresis. 2004 May;25(9):1327-33
pubmed: 15174055
J Mol Biol. 1998 Jun 26;279(5):1045-51
pubmed: 9642082
Mol Microbiol. 2018 Feb;107(4):445-454
pubmed: 29235176
Biochemistry. 2009 Nov 17;48(45):10793-802
pubmed: 19824612
J Bacteriol. 2000 May;182(10):2741-5
pubmed: 10781541
Mol Genet Genomics. 2001 Oct;266(2):313-7
pubmed: 11683274
Proc Natl Acad Sci U S A. 1968 Mar;59(3):777-84
pubmed: 4868216
J Biol Chem. 1973 Aug 25;248(16):5720-30
pubmed: 4579428
RNA Biol. 2012 Oct;9(10):1288-301
pubmed: 22995830
Front Microbiol. 2020 Jan 09;10:2982
pubmed: 31998269
Translation (Austin). 2013 Apr 01;1(1):e24315
pubmed: 26824016
Mol Cell. 2013 Nov 21;52(4):506-16
pubmed: 24207057
Acta Crystallogr D Biol Crystallogr. 2013 Jun;69(Pt 6):925-33
pubmed: 23695237
RNA. 2013 Jun;19(6):789-802
pubmed: 23611982
Sci Rep. 2015 Feb 25;5:8573
pubmed: 25712150
Nature. 2005 Dec 1;438(7068):628-32
pubmed: 16319883
J Mol Biol. 2013 Feb 22;425(4):767-79
pubmed: 23228329
Cell. 2002 Oct 4;111(1):129-40
pubmed: 12372306
Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):980-985
pubmed: 28096346
Methods Enzymol. 2013;530:159-72
pubmed: 24034320
Science. 2000 Aug 11;289(5481):905-20
pubmed: 10937989
Proc Natl Acad Sci U S A. 2015 Sep 1;112(35):10944-9
pubmed: 26283392
Elife. 2014 Oct 14;3:
pubmed: 25313868
Mol Cell Proteomics. 2002 May;1(5):376-86
pubmed: 12118079
J Biol Chem. 2018 Nov 30;293(48):18702-18709
pubmed: 30108177
Mol Microbiol. 1995 Aug;17(3):523-31
pubmed: 8559071
Nature. 2009 Feb 19;457(7232):977-80
pubmed: 19225518
J Biol Chem. 2018 Nov 30;293(48):18719-18729
pubmed: 30323064
J Bacteriol. 2008 Sep;190(17):5944-52
pubmed: 18621905
Nucleic Acids Res. 2016 Nov 16;44(20):9918-9932
pubmed: 27382067
Philos Trans R Soc Lond B Biol Sci. 2017 Mar 19;372(1716):
pubmed: 28138067