#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

A Conserved Pattern of Primer-Dependent Transcription Initiation in and Revealed by 5′ RNA-seq


Primer-dependent transcription initiation, PDI, refers to an alternative mechanism of transcription initiation whereby the first phosphodiester bond within the nascent RNA is formed between a 2- to ~4-nt RNA primer and an incoming nucleoside triphosphate. Although PDI has been shown to occur in E. coli, the impact of PDI on E. coli physiology, and the extent to which PDI occurs in other bacteria is unknown. Here we establish that PDI modulates the ability of E. coli to form biofilms, a surface attached community of bacteria encased in a polymeric matrix. We further describe a significantly improved RNA-seq based method for the detection of PDI in cells. Using this method we document the occurrence of PDI in the pathogenic bacterium Vibrio cholerae. We further show that the pattern of PDI in V. cholerae is identical to that observed in E. coli, suggesting that PDI in these two organisms may occur through a conserved process that produces identical populations of 2- to ~4-nt RNA primers. Our findings suggest PDI may be widespread in Gammaproteobacteria.


Vyšlo v časopise: A Conserved Pattern of Primer-Dependent Transcription Initiation in and Revealed by 5′ RNA-seq. PLoS Genet 11(7): e32767. doi:10.1371/journal.pgen.1005348
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1005348

Souhrn

Primer-dependent transcription initiation, PDI, refers to an alternative mechanism of transcription initiation whereby the first phosphodiester bond within the nascent RNA is formed between a 2- to ~4-nt RNA primer and an incoming nucleoside triphosphate. Although PDI has been shown to occur in E. coli, the impact of PDI on E. coli physiology, and the extent to which PDI occurs in other bacteria is unknown. Here we establish that PDI modulates the ability of E. coli to form biofilms, a surface attached community of bacteria encased in a polymeric matrix. We further describe a significantly improved RNA-seq based method for the detection of PDI in cells. Using this method we document the occurrence of PDI in the pathogenic bacterium Vibrio cholerae. We further show that the pattern of PDI in V. cholerae is identical to that observed in E. coli, suggesting that PDI in these two organisms may occur through a conserved process that produces identical populations of 2- to ~4-nt RNA primers. Our findings suggest PDI may be widespread in Gammaproteobacteria.


Zdroje

1. Feklistov A. RNA polymerase: in search of promoters. Annals of the New York Academy of Sciences. 2013;1293:25–32. doi: 10.1111/nyas.12197 23855603

2. Grunberg S, Hahn S. Structural insights into transcription initiation by RNA polymerase II. Trends Biochem Sci. 2013;38(12):603–11. doi: 10.1016/j.tibs.2013.09.002 24120742

3. Nickels BE, Dove SL. NanoRNAs: a class of small RNAs that can prime transcription initiation in bacteria. J Mol Biol. 2011;412(5):772–81. Epub 2011/06/28. doi: 10.1016/j.jmb.2011.06.015 21704045

4. Nickels BE. A new way to start: nanoRNA-mediated priming of transcription initiation. Transcription. 2012;3(6):300–4. doi: 10.4161/trns.21903 23117822

5. Vvedenskaya IO, Sharp JS, Goldman SR, Kanabar PN, Livny J, Dove SL, et al. Growth phase-dependent control of transcription start site selection and gene expression by nanoRNAs. Genes Dev. 2012;26(13):1498–507. doi: 10.1101/gad.192732.112 22751503

6. Goldman SR, Sharp JS, Vvedenskaya IO, Livny J, Dove SL, Nickels BE. NanoRNAs prime transcription initiation in vivo. Mol Cell. 2011;42(6):817–25. Epub 2011/06/28. doi: 10.1016/j.molcel.2011.06.005 21700226

7. Ghosh S, Deutscher MP. Oligoribonuclease is an essential component of the mRNA decay pathway. Proc Natl Acad Sci U S A. 1999;96(8):4372–7. 10200269

8. Mechold U, Fang G, Ngo S, Ogryzko V, Danchin A. YtqI from Bacillus subtilis has both oligoribonuclease and pAp-phosphatase activity. Nucleic Acids Res. 2007;35(13):4552–61. 17586819

9. Fang M, Zeisberg WM, Condon C, Ogryzko V, Danchin A, Mechold U. Degradation of nanoRNA is performed by multiple redundant RNases in Bacillus subtilis. Nucleic Acids Res. 2009;37(15):5114–25. Epub 2009/06/26. doi: 10.1093/nar/gkp527 19553197

10. Hoffman DJ, Niyogi SK. RNA initiation with dinucleoside monophosphates during transcription of bacteriophage T4 DNA with RNA polymerase of Escherichia coli. Proc Natl Acad Sci U S A. 1973;70(2):574–8. Epub 1973/02/01. 4568732

11. Minkley EG, Pribnow D. Transcription of the early region of bacteriophage T7: selective initiation with dinucleotides. J Mol Biol. 1973;77(2):255–77. Epub 1973/06/25. 4587752

12. Di Nocera PP, Avitabile A, Blasi F. In vitro transcription of the Escherichia coli histidine operon primed by dinucleotides. Effect of the first histidine biosynthetic enzyme. J Biol Chem. 1975;250(21):8376–81. Epub 1975/11/10. 1104602

13. Smagowicz WJ, Scheit KH. Primed abortive initiation of RNA synthesis by E. coli RNA polymerase on T7 DNA. Steady state kinetic studies. Nucleic Acids Res. 1978;5(6):1919–32. Epub 1978/06/01. 353734

14. Grachev MA, Zaychikov EF, Ivanova EM, Komarova NI, Kutyavin IV, Sidelnikova NP, et al. Oligonudeotides complementary to a promoter over the region −8… + 2 as transcription primers for E. coli RNA polymerase. Nucleic Acids Res. 1984;12(22):8509–24. 6390344

15. Ruetsch N, Dennis D. RNA polymerase. Limit cognate primer for initiation and stable ternary complex formation. J Biol Chem. 1987;262(4):1674–9. 3543009

16. Vvedenskaya IO, Goldman SR, Nickels BE. Preparation of cDNA libraries for high-throughput RNA sequencing analysis of RNA 5' ends. Methods Mol Biol. 2015;1276:211–28. doi: 10.1007/978-1-4939-2392-2_12 25665566

17. Zhang XS, Garcia-Contreras R, Wood TK. YcfR (BhsA) influences Escherichia coli biofilm formation through stress response and surface hydrophobicity. J Bacteriol. 2007;189(8):3051–62. 17293424

18. O'Toole GA. Microtiter dish biofilm formation assay. Journal of visualized experiments: JoVE. 2011;(47).

19. Maitra U, Hurwitz H. The role of DNA in RNA synthesis, IX. Nucleoside triphosphate termini in RNA polymerase products. Proc Natl Acad Sci U S A. 1965;54(3):815–22. 5324397

20. Jorgensen SE, Buch LB, Nierlich DP. Nucleoside triphosphate termini from RNA synthesized in vivo by Escherichia coli. Science. 1969;164(3883):1067–70. 4890175

21. Hawley DK, McClure WR. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983;11(8):2237–55. 6344016

22. Munafo DB, Robb GB. Optimization of enzymatic reaction conditions for generating representative pools of cDNA from small RNA. RNA. 2010;16(12):2537–52. doi: 10.1261/rna.2242610 20921270

23. Hafner M, Renwick N, Brown M, Mihailovic A, Holoch D, Lin C, et al. RNA-ligase-dependent biases in miRNA representation in deep-sequenced small RNA cDNA libraries. RNA. 2011;17(9):1697–712. doi: 10.1261/rna.2799511 21775473

24. Jayaprakash AD, Jabado O, Brown BD, Sachidanandam R. Identification and remediation of biases in the activity of RNA ligases in small-RNA deep sequencing. Nucleic Acids Res. 2011;39(21):e141. doi: 10.1093/nar/gkr693 21890899

25. Zhuang F, Fuchs RT, Sun Z, Zheng Y, Robb GB. Structural bias in T4 RNA ligase-mediated 3'-adapter ligation. Nucleic Acids Res. 2012;40(7):e54. doi: 10.1093/nar/gkr1263 22241775

26. Camilli A, Bassler BL. Bacterial small-molecule signaling pathways. Science. 2006;311(5764):1113–6. 16497924

27. Hengge R. Principles of c-di-GMP signalling in bacteria. Nat Rev Microbiol. 2009;7(4):263–73. Epub 2009/03/17. doi: 10.1038/nrmicro2109 19287449

28. Schirmer T, Jenal U. Structural and mechanistic determinants of c-di-GMP signalling. Nat Rev Microbiol. 2009;7(10):724–35. Epub 2009/09/17. doi: 10.1038/nrmicro2203 19756011

29. Boyd CD, O'Toole GA. Second messenger regulation of biofilm formation: breakthroughs in understanding c-di-GMP effector systems. Annu Rev Cell Dev Biol. 2012;28:439–62. doi: 10.1146/annurev-cellbio-101011-155705 23057745

30. Sondermann H, Shikuma NJ, Yildiz FH. You've come a long way: c-di-GMP signaling. Curr Opin Microbiol. 2012;15(2):140–6. doi: 10.1016/j.mib.2011.12.008 22226607

31. Romling U, Amikam D. Cyclic di-GMP as a second messenger. Curr Opin Microbiol. 2006;9(2):218–28. Epub 2006/03/15. 16530465

32. Davies BW, Bogard RW, Young TS, Mekalanos JJ. Coordinated regulation of accessory genetic elements produces cyclic di-nucleotides for V. cholerae virulence. Cell. 2012;149(2):358–70. doi: 10.1016/j.cell.2012.01.053 22500802

33. Danilchanka O, Mekalanos JJ. Cyclic dinucleotides and the innate immune response. Cell. 2013;154(5):962–70. doi: 10.1016/j.cell.2013.08.014 23993090

34. Corrigan RM, Grundling A. Cyclic di-AMP: another second messenger enters the fray. Nat Rev Microbiol. 2013;11(8):513–24. doi: 10.1038/nrmicro3069 23812326

35. Cameron DE, Urbach JM, Mekalanos JJ. A defined transposon mutant library and its use in identifying motility genes in Vibrio cholerae. Proc Natl Acad Sci U S A. 2008;105(25):8736–41. doi: 10.1073/pnas.0803281105 18574146

36. Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, et al. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol. 2006;2:2006 0008.

37. Pickering BS, Lopilato JE, Smith DR, Watnick PI. The Transcription Factor Mlc Promotes Vibrio cholerae Biofilm Formation through Repression of Phosphotransferase System Components. J Bacteriol. 2014;196(13):2423–30. doi: 10.1128/JB.01639-14 24769694

38. Chen D, Patton JT. Reverse transcriptase adds nontemplated nucleotides to cDNAs during 5'-RACE and primer extension. BioTechniques. 2001;30(3):574–80, 82. 11252793

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2015 Číslo 7
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#