1887

Abstract

The . Typhi gene, which codes for the LysR-type transcriptional regulator LeuO, contains five forward promoters named P3, P1, P2, P5 and P4, and two reverse promoters, P6 and P7. The activity of the forward promoters was revealed by primer extension using gene reporter fusions in an . Typhi mutant strain. Likewise, the activity of the reverse promoters was revealed in an background. Derepression of the transcription of the chromosomal gene was confirmed by RT-PCR in the mutant. The P1 transcriptional reporter fusion, which contained only the major P1 promoter, had a lower expression in a mutant strain, indicating that the steady-state levels of the (p)ppGpp alarmone positively regulate it. In contrast, the P3, P5P4, P6 and P7 transcriptional fusions were derepressed in the background, indicating that the alarmone has a negative effect on their expression. Thus, the search for genetic regulators and environmental cues that would differentially derepress gene expression by antagonizing the action of the H-NS and Lrp nucleoid-associated proteins, or that would fine-tune the expression of the various promoters, will further our understanding of the significance that multiple promoters have in the control of LeuO expression.

Funding
This study was supported by the:
  • Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de México (MX) (Award IN200517)
    • Principle Award Recipient: CalvaEdmundo
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001418
2021-09-30
2021-10-24
Loading full text...

Full text loading...

References

  1. Hernández-Lucas I, Calva E. The coming of age of the LeuO regulator. Mol Microbiol 2012; 85:1026–1028 [View Article] [PubMed]
    [Google Scholar]
  2. Lawley TD, Chan K, Thompson LJ, Kim CC, Govoni GR et al. Genome-wide screen for Salmonella genes required for long-term systemic infection of the mouse. PLoS Pathog 2006; 2:e11 [View Article] [PubMed]
    [Google Scholar]
  3. Rodríguez-Morales O, Fernández-Mora M, Hernández-Lucas I, Vázquez A, Puente JL et al. Salmonella enterica serovar Typhimurium ompS1 and ompS2 mutants are attenuated for virulence in mice. Infect Immun 2006; 74:1398–1402 [View Article] [PubMed]
    [Google Scholar]
  4. Tenor JL, McCormick BA, Ausubel FM, Aballay A. Caenorhabditis elegans-based screen identifies Salmonella virulence factors required for conserved host-pathogen interactions. Curr Biol 2004; 14:1018–1024 [View Article] [PubMed]
    [Google Scholar]
  5. Guadarrama C, Villaseñor T, Calva E. The subtleties and contrasts of the LeuO regulator in Salmonella Typhi: implications in the immune response. Front Immunol 2014; 5:581 [View Article] [PubMed]
    [Google Scholar]
  6. Hernández-Lucas I, Gallego-Hernández AL, Encarnación S, Fernández-Mora M, Martínez-Batallar AG et al. The LysR-type transcriptional regulator LeuO controls expression of several genes in Salmonella enterica serovar Typhi. J Bacteriol 2008; 190:1658–1670 [View Article] [PubMed]
    [Google Scholar]
  7. Dorman CJ. H-NS, the genome sentinel. Nat Rev Microbiol 2007; 5:157–161 [View Article] [PubMed]
    [Google Scholar]
  8. Dorman CJ, Schumacher MA, Bush MJ, Brennan RG, Buttner MJ. When is a transcription factor a NAP. Curr Opin Microbiol 2020; 55:26–33 [View Article] [PubMed]
    [Google Scholar]
  9. Chen CC, Chou MY, Huang CH, Majumder A, Wu HY. A cis-spreading nucleoprotein filament is responsible for the gene silencing activity found in the promoter relay mechanism. J Biol Chem 2005; 280:5101–5112 [View Article] [PubMed]
    [Google Scholar]
  10. Klauck E, Bohringer J, Hengge-Aronis R. The LysR-like regulator LeuO in Escherichia coli is involved in the translational regulation of rpoS by affecting the expression of the small regulatory DsrA-RNA. Mol Microbiol 1997; 25:559–569 [View Article] [PubMed]
    [Google Scholar]
  11. Breddermann H, Schnetz K. Activation of leuO by LrhA in Escherichia coli. Mol Microbiol 2017; 104:664–676 [View Article] [PubMed]
    [Google Scholar]
  12. Fang M, Wu HY. A promoter relay mechanism for sequential gene activation. J Bacteriol 1998; 180:626–633 [View Article] [PubMed]
    [Google Scholar]
  13. Stratmann T, Ü P, Wurm R, Wagner R, Schnetz K. RcsB-BglJ activates the Escherichia coli leuO gene, encoding an H-NS antagonist and pleiotropic regulator of virulence determinants. Mol Microbiol 2012; 83:1109–1123 [View Article] [PubMed]
    [Google Scholar]
  14. De la Cruz MA, Fernández-Mora M, Guadarrama C, Flores-Valdez MA, Bustamante VH et al. LeuO antagonizes H-NS and StpA-dependent repression in Salmonella enterica ompS1. Mol Microbiol 2007; 66:727–743 [View Article] [PubMed]
    [Google Scholar]
  15. Fernández-Mora M, Puente JL, Calva E. OmpR and LeuO positively regulate the Salmonella enterica serovar Typhi ompS2 porin gene. J Bacteriol 2004; 186:2909–2920 [View Article] [PubMed]
    [Google Scholar]
  16. Shimada T, Bridier A, Briandet R, Ishihama A. Novel roles of LeuO in transcription regulation of E. coli genome: antagonistic interplay with the universal silencer H-NS. Mol Microbiol 2011; 82:378–397 [View Article] [PubMed]
    [Google Scholar]
  17. Shimada T, Yamamoto K, Ishihama A. Involvement of the leucine response transcription factor LeuO in regulation of the genes for sulfa drug efflux. J Bacteriol 2009; 191:4562–4571 [View Article] [PubMed]
    [Google Scholar]
  18. Stratmann T, Madhusudan S, Schnetz K. Regulation of the yjjQ-bglJ operon, encoding LuxR-type transcription factors, and the divergent yjjP gene by H-NS and LeuO. J Bacteriol 2008; 190:926–935 [View Article] [PubMed]
    [Google Scholar]
  19. Fang M, Majumder A, Tsai KJ, Wu HY. ppGpp-dependent leuO expression in bacteria under stress. Biochem Biophys Res Commun 2000; 276:64–70 [View Article] [PubMed]
    [Google Scholar]
  20. Majumder A, Fang M, Tsai KJ, Ueguchi C, Mizuno T et al. LeuO expression in response to starvation for branched-chain amino acids. J Biol Chem 2001; 276:19046–19051 [View Article] [PubMed]
    [Google Scholar]
  21. VanBogelen RA, Olson ER, Wanner BL, Neidhardt FC. Global analysis of proteins synthesized during phosphorus restriction in Escherichia coli. J Bacteriol 1996; 178:4344–4366 [View Article] [PubMed]
    [Google Scholar]
  22. Ante VM, Bina XR, Bina JE. The LysR-type regulator LeuO regulates the acid tolerance response in Vibrio cholerae. Microbiology (Reading) 2015; 161:2434–2443 [View Article] [PubMed]
    [Google Scholar]
  23. Chen CC, Fang M, Majumder A, Wu HY. A 72-base pair AT-rich DNA sequence element functions as a bacterial gene silencer. J Biol Chem 2001; 276:9478–9485 [View Article] [PubMed]
    [Google Scholar]
  24. Dillon SC, Espinosa E, Hokamp K, Ussery DW, Casadesus J et al. LeuO is a global regulator of gene expression in Salmonella enterica serovar Typhimurium. Mol Microbiol 2012; 85:1072–1089 [View Article] [PubMed]
    [Google Scholar]
  25. Espinosa E, Casadesús J. Regulation of Salmonella enterica pathogenicity island (SPI-1) by the LysR-type regulator LeuO. Mol Microbiol 2014; 91:1057–1069 [View Article] [PubMed]
    [Google Scholar]
  26. Gallego-Hernández AL, Hernández-Lucas I, De la Cruz MA, Olvera L, Morett E et al. Transcriptional regulation of the assT-dsbL-dsbI gene cluster in Salmonella enterica serovar Typhi IMSS-1 depends on LeuO, H-NS, and specific growth conditions. J Bacteriol 2012; 194:2254–2264 [View Article] [PubMed]
    [Google Scholar]
  27. Lawrenz MB, Miller VL. Comparative analysis of the regulation of rovA from the pathogenic Yersiniae. J Bacteriol 2007; 189:5963–5975 [View Article] [PubMed]
    [Google Scholar]
  28. Medina-Aparicio L, Rebollar-Flores JE, Gallego-Hernández AL, Vázquez A, Olvera L et al. The CRISPR/Cas immune system is an operon regulated by LeuO, H-NS, and leucine-responsive regulatory protein in Salmonella enterica serovar Typhi. J Bacteriol 2011; 193:2396–2407 [View Article] [PubMed]
    [Google Scholar]
  29. Shi X, Bennett GN. Effects of multicopy LeuO on the expression of the acid-inducible lysine decarboxylase gene in Escherichia coli. J Bacteriol 1995; 177:810–814 [View Article] [PubMed]
    [Google Scholar]
  30. Ueguchi C, Ohta T, Seto C, Suzuki T, Mizuno T. The leuO gene product has a latent ability to relieve bgl silencing in Escherichia coli. J Bacteriol 1998; 180:190–193 [View Article] [PubMed]
    [Google Scholar]
  31. Westra ER, Pul U, Heidrich N, Jore MM, Lundgren M et al. H-NS-mediated repression of CRISPR-based immunity in Escherichia coli K12 can be relieved by the transcription activator LeuO. Mol Microbiol 2010; 77:1380–1393 [View Article] [PubMed]
    [Google Scholar]
  32. Puente JL, Flores V, Fernández M, Fuchs Y, Calva E. Isolation of an ompC-like outer membrane protein gene from Salmonella typhi. Gene 1987; 61:75–83 [View Article] [PubMed]
    [Google Scholar]
  33. Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 2000; 97:6640–6645 [View Article] [PubMed]
    [Google Scholar]
  34. Flores-Valdez MA, Puente JL, Calva E. Negative osmoregulation of the Salmonella ompS1 porin gene independently of OmpR in an hns background. J Bacteriol 2003; 185:6497–6506 [View Article] [PubMed]
    [Google Scholar]
  35. Shapira SK, Chou J, Richaud FV, Casadaban MJ. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene 1983; 25:71–82 [View Article] [PubMed]
    [Google Scholar]
  36. Simons RW, Houman F, Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene 1987; 53:85–96 [View Article] [PubMed]
    [Google Scholar]
  37. Oropeza R, Sampieri CL, Puente JL, Calva E. Negative and positive regulation of the non-osmoregulated ompS1 porin gene in Salmonella typhi: a novel regulatory mechanism that involves OmpR. Mol Microbiol 1999; 32:243–252 [View Article] [PubMed]
    [Google Scholar]
  38. Chen CC, Wu HY. LeuO protein delimits the transcriptionally active and repressive domains on the bacterial chromosome. J Biol Chem 2005; 280:15111–15121 [View Article] [PubMed]
    [Google Scholar]
  39. Yamada M, Kabir MS, Tsunedomi R. Divergent promoter organization may be a preferred structure for gene control in Escherichia coli. J Mol Microbiol Biotechnol 2003; 6:206–210 [View Article] [PubMed]
    [Google Scholar]
  40. Landgraf JR, Wu J, Calvo JM. Effects of nutrition and growth rate on Lrp levels in Escherichia coli. J Bacteriol 1996; 178:6930–6936 [View Article] [PubMed]
    [Google Scholar]
  41. Platko JV, Willins DA, Calvo JM. The ilvIH operon of Escherichia coli is positively regulated. J Bacteriol 1990; 172:4563–4570 [View Article] [PubMed]
    [Google Scholar]
  42. Dorman CJ, Dorman MJ. Control of virulence gene transcription by indirect readout in Vibrio cholerae and Salmonella enterica serovar Typhimurium. Environ Microbiol 2017; 19:3834–3845 [View Article] [PubMed]
    [Google Scholar]
  43. Pul U, Wurm R, Lux B, Meltzer M, Menzel A et al. LRP and H-NS--cooperative partners for transcription regulation at Escherichia coli rRNA promoters. Mol Microbiol 2005; 58:864–876 [View Article] [PubMed]
    [Google Scholar]
  44. Takao M, Yen H, Tobe T. LeuO enhances butyrate-induced virulence expression through a positive regulatory loop in enterohaemorrhagic Escherichia coli. Mol Microbiol 2014; 93:1302–1313 [View Article] [PubMed]
    [Google Scholar]
  45. McPheeters DS, Christensen A, Young ET, Stormo G, Gold L. Translational regulation of expression of the bacteriophage T4 lysozyme gene. Nucleic Acids Res 1986; 14:5813–5826 [View Article] [PubMed]
    [Google Scholar]
  46. Flores-Valdez MA, Fernández-Mora M, Ares MA, Girón JA, Calva E et al. OmpR phosphorylation regulates ompS1 expression by differentially controlling the use of promoters. Microbiology (Reading) 2014; 160:733–741 [View Article] [PubMed]
    [Google Scholar]
  47. Musso RE, Di Lauro R, Adhya S, de Crombrugghe B. Dual control for transcription of the galactose operon by cyclic AMP and its receptor protein at two interspersed promoters. Cell 1977; 12:847–854 [View Article] [PubMed]
    [Google Scholar]
  48. Nakagawa A, Oshima T, Mori H. Identification and characterization of a second, inducible promoter of relA in Escherichia coli. Genes Genet Syst 2006; 81:299–310 [View Article] [PubMed]
    [Google Scholar]
  49. Epshtein V, Nudler E. Cooperation between RNA polymerase molecules in transcription elongation. Science 2003; 300:801–805 [View Article] [PubMed]
    [Google Scholar]
  50. Epshtein V, Toulme F, Rahmouni AR, Borukhov S, Nudler E. Transcription through the roadblocks: the role of RNA polymerase cooperation. EMBO J 2003; 22:4719–4727 [View Article] [PubMed]
    [Google Scholar]
  51. Jia J, King JE, Goldrick MC, Aldawood E, Roberts IS. Three tandem promoters, together with IHF, regulate growth phase dependent expression of the Escherichia coli kps capsule gene cluster. Sci Rep 2017; 7:17924 [View Article] [PubMed]
    [Google Scholar]
  52. Richardson SM, Higgins CF, Lilley DM. DNA supercoiling and the leu-500 promoter mutation of Salmonella typhimurium. EMBO J 1988; 7:1863–1869 [View Article] [PubMed]
    [Google Scholar]
  53. Imholz NCE, Noga MJ, van den Broek NJF, Bokinsky G. Calibrating the bacterial growth rate speedometer: A re-evaluation of the relationship between basal ppGpp, growth, and RNA synthesis in Escherichia coli. Front Microbiol 2020; 11:574872 [View Article] [PubMed]
    [Google Scholar]
  54. Sánchez-Vázquez P, Dewey CN, Kitten N, Ross W, Gourse RL. Genome-wide effects on Escherichia coli transcription from ppGpp binding to its two sites on RNA polymerase. Proc Natl Acad Sci U S A 2019; 116:8310–8319 [View Article] [PubMed]
    [Google Scholar]
  55. Choi J, Groisman EA. Salmonella expresses foreign genes during infection by degrading their silencer. Proc Natl Acad Sci U S A 2020; 117:8074–8082 [View Article] [PubMed]
    [Google Scholar]
  56. Curran TD, Abacha F, Hibberd SP, Rolfe MD, Lacey MM et al. Identification of new members of the Escherichia coli K-12 MG1655 SlyA regulon. Microbiology (Reading) 2017; 163:400–409 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001418
Loading
/content/journal/jmm/10.1099/jmm.0.001418
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error