Small Molecule Inhibitors of RnpA Alter Cellular mRNA Turnover, Exhibit Antimicrobial Activity, and Attenuate Pathogenesis
Methicillin-resistant Staphylococcus aureus is estimated to cause more U.S. deaths annually than HIV/AIDS. The emergence of hypervirulent and multidrug-resistant strains has further amplified public health concern and accentuated the need for new classes of antibiotics. RNA degradation is a required cellular process that could be exploited for novel antimicrobial drug development. However, such discovery efforts have been hindered because components of the Gram-positive RNA turnover machinery are incompletely defined. In the current study we found that the essential S. aureus protein, RnpA, catalyzes rRNA and mRNA digestion in vitro. Exploiting this activity, high through-put and secondary screening assays identified a small molecule inhibitor of RnpA-mediated in vitro RNA degradation. This agent was shown to limit cellular mRNA degradation and exhibited antimicrobial activity against predominant methicillin-resistant S. aureus (MRSA) lineages circulating throughout the U.S., vancomycin intermediate susceptible S. aureus (VISA), vancomycin resistant S. aureus (VRSA) and other Gram-positive bacterial pathogens with high RnpA amino acid conservation. We also found that this RnpA-inhibitor ameliorates disease in a systemic mouse infection model and has antimicrobial activity against biofilm-associated S. aureus. Taken together, these findings indicate that RnpA, either alone, as a component of the RNase P holoenzyme, and/or as a member of a more elaborate complex, may play a role in S. aureus RNA degradation and provide proof of principle for RNA catabolism-based antimicrobial therapy.
Vyšlo v časopise:
Small Molecule Inhibitors of RnpA Alter Cellular mRNA Turnover, Exhibit Antimicrobial Activity, and Attenuate Pathogenesis. PLoS Pathog 7(2): e32767. doi:10.1371/journal.ppat.1001287
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1001287
Souhrn
Methicillin-resistant Staphylococcus aureus is estimated to cause more U.S. deaths annually than HIV/AIDS. The emergence of hypervirulent and multidrug-resistant strains has further amplified public health concern and accentuated the need for new classes of antibiotics. RNA degradation is a required cellular process that could be exploited for novel antimicrobial drug development. However, such discovery efforts have been hindered because components of the Gram-positive RNA turnover machinery are incompletely defined. In the current study we found that the essential S. aureus protein, RnpA, catalyzes rRNA and mRNA digestion in vitro. Exploiting this activity, high through-put and secondary screening assays identified a small molecule inhibitor of RnpA-mediated in vitro RNA degradation. This agent was shown to limit cellular mRNA degradation and exhibited antimicrobial activity against predominant methicillin-resistant S. aureus (MRSA) lineages circulating throughout the U.S., vancomycin intermediate susceptible S. aureus (VISA), vancomycin resistant S. aureus (VRSA) and other Gram-positive bacterial pathogens with high RnpA amino acid conservation. We also found that this RnpA-inhibitor ameliorates disease in a systemic mouse infection model and has antimicrobial activity against biofilm-associated S. aureus. Taken together, these findings indicate that RnpA, either alone, as a component of the RNase P holoenzyme, and/or as a member of a more elaborate complex, may play a role in S. aureus RNA degradation and provide proof of principle for RNA catabolism-based antimicrobial therapy.
Zdroje
1. ShorrAF
TabakYP
KillianAD
GuptaV
LiuLZ
2006 Healthcare-associated bloodstream infection: A distinct entity? Insights from a large U.S. database. Crit Care Med 34 2588 2595
2. BancroftEA
2007 Antimicrobial resistance: it's not just for hospitals. Jama 298 1803 1804
3. KlevensRM
MorrisonMA
NadleJ
PetitS
GershmanK
2007 Invasive methicillin-resistant Staphylococcus aureus infections in the United States. Jama 298 1763 1771
4. AppelbaumPC
2007 Reduced glycopeptide susceptibility in methicillin-resistant Staphylococcus aureus (MRSA). Int J Antimicrob Agents 30 398 408
5. ZetolaN
FrancisJS
NuermbergerEL
BishaiWR
2005 Community-acquired meticillin-resistant Staphylococcus aureus: an emerging threat. Lancet Infect Dis 5 275 286
6. CarpousisAJ
2007 The RNA degradosome of Escherichia coli: an mRNA-degrading machine assembled on RNase E. Annu Rev Microbiol 61 71 87
7. MackieGA
1998 Ribonuclease E is a 5′-end-dependent endonuclease. Nature 395 720 723
8. VanzoNF
LiYS
PyB
BlumE
HigginsCF
1998 Ribonuclease E organizes the protein interactions in the Escherichia coli RNA degradosome. Genes Dev 12 2770 2781
9. CondonC
2003 RNA processing and degradation in Bacillus subtilis. Microbiol Mol Biol Rev 67 157 174, table of contents
10. EvenS
PellegriniO
ZigL
LabasV
VinhJ
2005 Ribonucleases J1 and J2: two novel endoribonucleases in B. subtilis with functional homology to E. coli RNase E. Nucleic Acids Res 33 2141 2152. Print 2005
11. MathyN
BenardL
PellegriniO
DaouR
WenT
2007 5′-to-3′ exoribonuclease activity in bacteria: role of RNase J1 in rRNA maturation and 5′ stability of mRNA. Cell 129 681 692
12. CommichauFM
RotheFM
HerzbergC
WagnerE
HellwigD
2009 Novel activities of glycolytic enzymes in Bacillus subtilis: interactions with essential proteins involved in mRNA processing. Mol Cell Proteomics 8 1350 1360
13. MaderU
ZigL
KretschmerJ
HomuthG
PutzerH
2008 mRNA processing by RNases J1 and J2 affects Bacillus subtilis gene expression on a global scale. Mol Microbiol 70 183 196
14. YaoS
SharpJS
BechhoferDH
2009 Bacillus subtilis RNase J1 endonuclease and 5′ exonuclease activities in the turnover of DeltaermC mRNA. Rna 15 2331 2339
15. ShahbabianK
JamalliA
ZigL
PutzerH
2009 RNase Y, a novel endoribonuclease, initiates riboswitch turnover in Bacillus subtilis. Embo J 28 3523 3533
16. Lehnik-HabrinkM
PfortnerH
RempetersL
PietackN
HerzbergC
2010 The RNA degradosome in Bacillus subtilis: identification of CshA as the major RNA helicase in the multiprotein complex. Mol Microbiol 21 21
17. KobayashiK
EhrlichSD
AlbertiniA
AmatiG
AndersenKK
2003 Essential Bacillus subtilis genes. Proc Natl Acad Sci U S A 100 4678 4683
18. NovickRP
2003 Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol Microbiol 48 1429 1449
19. AndersonKL
RobertsC
DiszT
VonsteinV
HwangK
2006 Characterization of the Staphylococcus aureus heat-shock, cold-shock, stringent, and SOS responses and their effects on log-phase mRNA turnover. J Bacteriol 188 6739 6756
20. RobertsC
AndersonKL
MurphyE
ProjanSJ
MountsW
2006 Characterizing the Effect of the Staphylococcus aureus Virulence Factor Regulator, SarA, on Log-Phase mRNA Half-Lives. J Bacteriol 188 2593 2603
21. TakayamaK
KjellebergS
2000 The role of RNA stability during bacterial stress responses and starvation. Environ Microbiol 2 355 365
22. AndersonKL
DunmanPM
2009 Messenger RNA Turnover Processes in Escherichia coli, Bacillus subtilis, and Emerging Studies in Staphylococcus aureus. Int J Microbiol 2009 525491
23. FrankDN
PaceNR
1998 Ribonuclease P: unity and diversity in a tRNA processing ribozyme. Annu Rev Biochem 67 153 180
24. KazantsevAV
PaceNR
2006 Bacterial RNase P: a new view of an ancient enzyme. Nat Rev Microbiol 4 729 740
25. WalkerSC
EngelkeDR
2006 Ribonuclease P: the evolution of an ancient RNA enzyme. Crit Rev Biochem Mol Biol 41 77 102
26. HartmannRK
GossringerM
SpathB
FischerS
MarchfelderA
2009 The making of tRNAs and more - RNase P and tRNase Z. Prog Mol Biol Transl Sci 85 319 368
27. GossringerM
Kretschmer-Kazemi FarR
HartmannRK
2006 Analysis of RNase P protein (rnpA) expression in Bacillus subtilis utilizing strains with suppressible rnpA expression. J Bacteriol 188 6816 6823
28. SchedlP
PrimakoffP
1973 Mutants of Escherichia coli thermosensitive for the synthesis of transfer RNA. Proc Natl Acad Sci U S A 70 2091 2095
29. WaughDS
PaceNR
1990 Complementation of an RNase P RNA (rnpB) gene deletion in Escherichia coli by homologous genes from distantly related eubacteria. J Bacteriol 172 6316 6322
30. LundbladEW
XiaoG
KoJH
AltmanS
2008 Rapid selection of accessible and cleavable sites in RNA by Escherichia coli RNase P and random external guide sequences. Proc Natl Acad Sci U S A 105 2354 2357
31. LiuF
AltmanS
1994 Differential evolution of substrates for an RNA enzyme in the presence and absence of its protein cofactor. Cell 77 1093 1100
32. MarvinMC
EngelkeDR
2009 Broadening the mission of an RNA enzyme. J Cell Biochem 108 1244 1251
33. LetunicI
CopleyRR
PilsB
PinkertS
SchultzJ
2006 SMART 5: domains in the context of genomes and networks. Nucleic Acids Res 34 D257 260
34. SchultzJ
MilpetzF
BorkP
PontingCP
1998 SMART, a simple modular architecture research tool: identification of signaling domains. Proc Natl Acad Sci U S A 95 5857 5864
35. SpitzfadenC
NicholsonN
JonesJJ
GuthS
LehrR
2000 The structure of ribonuclease P protein from Staphylococcus aureus reveals a unique binding site for single-stranded RNA. J Mol Biol 295 105 115
36. ChaudhuriRR
AllenAG
OwenPJ
ShalomG
StoneK
2009 Comprehensive identification of essential Staphylococcus aureus genes using Transposon-Mediated Differential Hybridisation (TMDH). BMC Genomics 10 291
37. JiY
ZhangB
VanSF
Horn
WarrenP
2001 Identification of critical staphylococcal genes using conditional phenotypes generated by antisense RNA. Science 293 2266 2269
38. CharpentierE
AntonAI
BarryP
AlfonsoB
FangY
2004 Novel cassette-based shuttle vector system for gram-positive bacteria. Appl Environ Microbiol 70 6076 6085
39. McDougalLK
StewardCD
KillgoreGE
ChaitramJM
McAllisterSK
2003 Pulsed-field gel electrophoresis typing of oxacillin-resistant Staphylococcus aureus isolates from the United States: establishing a national database. J Clin Microbiol 41 5113 5120
40. WeissEC
SpencerHJ
DailySJ
WeissBD
SmeltzerMS
2009 Impact of sarA on antibiotic susceptibility of Staphylococcus aureus in a catheter-associated in vitro model of biofilm formation. Antimicrob Agents Chemother 53 2475 2482
41. BeenkenKE
DunmanPM
McAleeseF
MacapagalD
MurphyE
2004 Global Gene Expression in Staphylococcus aureus Biofilms. J Bacteriol 186 4665 4684
42. HuntzingerE
BoissetS
SaveanuC
BenitoY
GeissmannT
2005 Staphylococcus aureus RNAIII and the endoribonuclease III coordinately regulate spa gene expression. EMBO J 24 824 835. Epub 2005 Jan 2027
43. KoleR
BaerMF
StarkBC
AltmanS
1980 E. coli RNAase P has a required RNA component. Cell 19 881 887
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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