Molecular Characterization of Global Regulatory RNA Species That Control Pathogenicity Factors in Erwinia Amylovora and Erwinia Herbicola Pv. Gypsophilae

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2001 Feb 27

PMID 11222584

Citations 17

Authors

W Ma

Y Cui

Y Liu

C K Dumenyo

A Mukherjee

A K Chatterjee

Affiliations

Soon will be listed here.

Abstract

rsmB(Ecc) specifies a nontranslatable RNA regulator that controls exoprotein production and pathogenicity in soft rot-causing Erwinia carotovora subsp. carotovora. This effect of rsmB(Ecc) RNA is mediated mostly by neutralizing the function of RsmA(Ecc), an RNA-binding protein of E. carotovora subsp. carotovora, which acts as a global negative regulator. To determine the occurrence of functional homologs of rsmB(Ecc) in non-soft-rot-causing Erwinia species, we cloned the rsmB genes of E. amylovora (rsmB(Ea)) and E. herbicola pv. gypsophilae (rsmB(Ehg)). We show that rsmB(Ea) in E. amylovora positively regulates extracellular polysaccharide (EPS) production, motility, and pathogenicity. In E. herbicola pv. gypsophilae, rsmB(Ehg) elevates the levels of transcripts of a cytokinin (etz) gene and stimulates the production of EPS and yellow pigment as well as motility. RsmA(Ea) and RsmA(Ehg) have more than 93% identity to RsmA(Ecc) and, like the latter, function as negative regulators by affecting the transcript stability of the target gene. The rsmB genes reverse the negative effects of RsmA(Ea), RsmA(Ehg), and RsmA(Ecc), but the extent of reversal is highest with homologous combinations of rsm genes. These observations and findings that rsmB(Ea) and rsmB(Ehg) RNA bind RsmA(Ecc) indicate that the rsmB effect is channeled via RsmA. Additional support for this conclusion comes from the observation that the rsmB genes are much more effective as positive regulators in a RsmA(+) strain of E. carotovora subsp. carotovora than in its RsmA(-) derivative. E. herbicola pv. gypsophilae produces a 290-base rsmB transcript that is not subject to processing. By contrast, E. amylovora produces 430- and 300-base rsmB transcripts, the latter presumably derived by processing of the primary transcript as previously noted with the transcripts of rsmB(Ecc). Southern blot hybridizations revealed the presence of rsmB homologs in E. carotovora, E. chrysanthemi, E. amylovora, E. herbicola, E. stewartii and E. rhapontici, as well as in other enterobacteria such as Escherichia coli, Salmonella enterica serovar Typhimurium, Serratia marcescens, Shigella flexneri, Enterobacter aerogenes, Klebsiella pneumoniae, Yersinia enterocolitica, and Y. pseudotuberculosis. A comparison of rsmB sequences from several of these enterobacterial species revealed a highly conserved 34-mer region which is predicted to play a role in positive regulation by rsmB RNA.

Citing Articles

The rsmS (ybaM) mutation causes bypass suppression of the RsmAB post-transcriptional virulence regulation system in enterobacterial phytopathogens.

Monson R, Apagyi K, Bowden S, Simpson N, Williamson N, Cubitt M Sci Rep. 2019; 9(1):4525.

PMID: 30872786 PMC: 6418279. DOI: 10.1038/s41598-019-40970-3.

Comparative transcriptome analysis of a lowly virulent strain of Erwinia amylovora in shoots of two apple cultivars - susceptible and resistant to fire blight.

Pulawska J, Kaluzna M, Warabieda W, Mikicinski A BMC Genomics. 2017; 18(1):868.

PMID: 29132313 PMC: 5683332. DOI: 10.1186/s12864-017-4251-z.

CsrB, a noncoding regulatory RNA, is required for BarA-dependent expression of biocontrol traits in Rahnella aquatilis HX2.

Mei L, Xu S, Lu P, Lin H, Guo Y, Wang Y PLoS One. 2017; 12(11):e0187492.

PMID: 29091941 PMC: 5665550. DOI: 10.1371/journal.pone.0187492.

Structural rearrangement in an RsmA/CsrA ortholog of Pseudomonas aeruginosa creates a dimeric RNA-binding protein, RsmN.

Morris E, Hall G, Li C, Heeb S, Kulkarni R, Lovelock L Structure. 2013; 21(9):1659-71.

PMID: 23954502 PMC: 3791407. DOI: 10.1016/j.str.2013.07.007.

Discovery of plant phenolic compounds that act as type III secretion system inhibitors or inducers of the fire blight pathogen, Erwinia amylovora.

Khokhani D, Zhang C, Li Y, Wang Q, Zeng Q, Yamazaki A Appl Environ Microbiol. 2013; 79(18):5424-36.

PMID: 23770912 PMC: 3754148. DOI: 10.1128/AEM.00845-13.

References

Chatterjee A, McEvoy J, Chambost J, Blasco F, Chatterjee A . Nucleotide sequence and molecular characterization of pnlA, the structural gene for damage-inducible pectin lyase of Erwinia carotovora subsp. carotovora 71. J Bacteriol. 1991; 173(5):1765-9. PMC: 207328. DOI: 10.1128/jb.173.5.1765-1769.1991. View

Bugert P, Geider K . Molecular analysis of the ams operon required for exopolysaccharide synthesis of Erwinia amylovora. Mol Microbiol. 1995; 15(5):917-33. DOI: 10.1111/j.1365-2958.1995.tb02361.x. View

Mukherjee A, Cui Y, Liu Y, Dumenyo C, Chatterjee A . Global regulation in Erwinia species by Erwinia carotovora rsmA, a homologue of Escherichia coli csrA: repression of secondary metabolites, pathogenicity and hypersensitive reaction. Microbiology (Reading). 1996; 142 ( Pt 2):427-434. DOI: 10.1099/13500872-142-2-427. View

Hauben L, Moore E, Vauterin L, Steenackers M, Mergaert J, Verdonck L . Phylogenetic position of phytopathogens within the Enterobacteriaceae. Syst Appl Microbiol. 1998; 21(3):384-97. DOI: 10.1016/S0723-2020(98)80048-9. View

Aiba H, Adhya S, de Crombrugghe B . Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981; 256(22):11905-10. View

White D, Hart M, Romeo T . Phylogenetic distribution of the global regulatory gene csrA among eubacteria. Gene. 1996; 182(1-2):221-3. DOI: 10.1016/s0378-1119(96)00547-1. View

Cui Y, Chatterjee A, Liu Y, Dumenyo C, Chatterjee A . Identification of a global repressor gene, rsmA, of Erwinia carotovora subsp. carotovora that controls extracellular enzymes, N-(3-oxohexanoyl)-L-homoserine lactone, and pathogenicity in soft-rotting Erwinia spp. J Bacteriol. 1995; 177(17):5108-15. PMC: 177290. DOI: 10.1128/jb.177.17.5108-5115.1995. View

Selvaraj G, Fong Y, Iyer V . A portable DNA sequence carrying the cohesive site (cos) of bacteriophage lambda and the mob (mobilization) region of the broad-host-range plasmid RK2: a module for the construction of new cosmids. Gene. 1984; 32(1-2):235-41. DOI: 10.1016/0378-1119(84)90051-9. View

Mukherjee A, Cui Y, Ma W, Liu Y, Chatterjee A . hexA of Erwinia carotovora ssp. carotovora strain Ecc71 negatively regulates production of RpoS and rsmB RNA, a global regulator of extracellular proteins, plant virulence and the quorum-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone. Environ Microbiol. 2001; 2(2):203-15. DOI: 10.1046/j.1462-2920.2000.00093.x. View

10.

Liu Y, Murata H, Chatterjee A, Chatterjee A . Characterization of a novel regulatory gene aepA that controls extracellular enzyme production in the phytopathogenic bacterium Erwinia carotovora subsp. carotovora. Mol Plant Microbe Interact. 1993; 6(3):299-308. DOI: 10.1094/mpmi-6-299. View

11.

Barras F, Thurn K, Chatterjee A . Resolution of four pectate lyase structural genes of Erwinia chrysanthemi (EC16) and characterization of the enzymes produced in Escherichia coli. Mol Gen Genet. 1987; 209(2):319-25. DOI: 10.1007/BF00329660. View

12.

Chatterjee A, Thurn K, TYRELL D . Isolation and characterization of Tn5 insertion mutants of Erwinia chrysanthemi that are deficient in polygalacturonate catabolic enzymes oligogalacturonate lyase and 3-deoxy-D-glycero-2,5-hexodiulosonate dehydrogenase. J Bacteriol. 1985; 162(2):708-14. PMC: 218908. DOI: 10.1128/jb.162.2.708-714.1985. View

13.

Fleischmann R, Adams M, White O, Clayton R, Kirkness E, Kerlavage A . Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995; 269(5223):496-512. DOI: 10.1126/science.7542800. View

14.

Zink R, Kemble R, Chatterjee A . Transposon Tn5 mutagenesis in Erwinia carotovora subsp. carotovora and E. carotovora subsp. atroseptica. J Bacteriol. 1984; 157(3):809-14. PMC: 215331. DOI: 10.1128/jb.157.3.809-814.1984. View

15.

Cotter P, Miller J . In vivo and ex vivo regulation of bacterial virulence gene expression. Curr Opin Microbiol. 1999; 1(1):17-26. DOI: 10.1016/s1369-5274(98)80138-0. View

16.

Keen N, Tamaki S, Kobayashi D, Trollinger D . Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988; 70(1):191-7. DOI: 10.1016/0378-1119(88)90117-5. View

17.

Manulis S, Brandl M, Lindow S, Barash I . Differential involvement of indole-3-acetic acid biosynthetic pathways in pathogenicity and epiphytic fitness of Erwinia herbicola pv. gypsophilae. Mol Plant Microbe Interact. 1998; 11(7):634-42. DOI: 10.1094/MPMI.1998.11.7.634. View

18.

Lerner C, Inouye M . Low copy number plasmids for regulated low-level expression of cloned genes in Escherichia coli with blue/white insert screening capability. Nucleic Acids Res. 1990; 18(15):4631. PMC: 331321. DOI: 10.1093/nar/18.15.4631. View

19.

Mukherjee A, Cui Y, Liu Y, Chatterjee A . Molecular characterization and expression of the Erwinia carotovora hrpNEcc gene, which encodes an elicitor of the hypersensitive reaction. Mol Plant Microbe Interact. 1997; 10(4):462-71. DOI: 10.1094/MPMI.1997.10.4.462. View

20.

McEvoy J, Murata H, Chatterjee A . Molecular cloning and characterization of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents. J Bacteriol. 1990; 172(6):3284-9. PMC: 209137. DOI: 10.1128/jb.172.6.3284-3289.1990. View