Identification of a Gene Involved in the Negative Regulation of Pyomelanin Production in Ralstonia solanacearum
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Ralstonia solanacearum causes bacterial wilt in a wide variety of host plant species and produces a melanin-like blackish-brown pigment in stationary phase when grown in minimal medium supplemented with tyrosine. To study melanin production regulation in R. solanacearum, five mutants exhibiting overproduction of melanin-like pigments were selected from a transposon (Tn) insertion mutant library of R. solanacearum SL341. Most of the mutants, except one (SL341T), were not complemented by the original gene or overproduced melanins. SL341T showed Tn insertion in a gene containing a conserved domain of eukaryotic transcription factor. The gene was annotated as a hypothetical protein, given its weak similarity to any known proteins. Upon complementation with its original gene, the mutant strains reverted to their wild-type phenotype. SL341T produced 3-folds more melanin at 72 h post-incubation compared with wild-type SL341 when grown in minimal medium supplemented with tyrosine. The chemical analysis of SL341T cultural filtrate revealed the accumulation of a higher amount of homogentisate, a major precursor of pyomelanin, and a lower amount of dihydroxyphenylalanine, an intermediate of eumelanin, compared with SL341. The expression study showed a relatively higher expression of hppD (encoding hydroxyphenylpyruvate dioxygenase) and lower expression of hmgA (encoding homogentisate dioxygenase) and nagL (encoding maleylacetoacetate isomerase) in SL341T than in SL341. SL341 showed a significantly higher expression of tyrosinase gene compared with SL341T at 48 h post-incubation. These results indicated that R. solanacearum produced both pyomelanin and eumelanin, and the novel hypothetical protein is involved in the negative regulation of melanin production.
Abstract
Ralstonia solanacearum causes bacterial wilt in a wide variety of host plant species and produces a melanin-like blackish-brown pigment in stationary phase when grown in minimal medium supplemented with tyrosine. To study melanin production regulation in R. solanacearum, five mutants exhibiting overproduction of melanin-like pigments were selected from a transposon (Tn) insertion mutant library of R. solanacearum SL341. Most of the mutants, except one (SL341T), were not complemented by the original gene or overproduced melanins. SL341T showed Tn insertion in a gene containing a conserved domain of eukaryotic transcription factor. The gene was annotated as a hypothetical protein, given its weak similarity to any known proteins. Upon complementation with its original gene, the mutant strains reverted to their wild-type phenotype. SL341T produced 3-folds more melanin at 72 h post-incubation compared with wild-type SL341 when grown in minimal medium supplemented with tyrosine. The chemical analysis of SL341T cultural filtrate revealed the accumulation of a higher amount of homogentisate, a major precursor of pyomelanin, and a lower amount of dihydroxyphenylalanine, an intermediate of eumelanin, compared with SL341. The expression study showed a relatively higher expression of hppD (encoding hydroxyphenylpyruvate dioxygenase) and lower expression of hmgA (encoding homogentisate dioxygenase) and nagL (encoding maleylacetoacetate isomerase) in SL341T than in SL341. SL341 showed a significantly higher expression of tyrosinase gene compared with SL341T at 48 h post-incubation. These results indicated that R. solanacearum produced both pyomelanin and eumelanin, and the novel hypothetical protein is involved in the negative regulation of melanin production.
참고문헌 (39)
- Riley PA., "Melanin", Int. J. Biochem. Cell Biol, 29, pp.1235~1239 (1997)
- Nosanchuk JD, "Cellular charge of Cryptococcus neoformans: contributions from the capsular polysaccharide, melanin, and monoclonal antibody binding", Infect. Immun, 65, pp.1836~1841 (1997)
- White LP, "Melanin: a naturally occurring cation exchange material", Nature, 182, pp.1427~1428 (1958)
- Nosanchuk JD, "The contribution of melanin to microbial pathogenesis", Cell. Microbiol, 5, pp.203~223 (2003)
- Hill HZ, "The function of melanin or six blind people examine an elephant", BioEssays, 14, pp.49~56 (1992)
- Fogarty RV, "Fungal melanins and their interactions with metals", Enzyme Microb. Technol, 19, pp.311~317 (1996)
- Han H, "Loss of homogentisate 1,2-dioxygenase activity in Bacillus anthracis results in accumulation of protective pigment", PLoS One, 10, pp.e0128967~ (2015)
- Rodríguez-Rojas A, "Inactivation of the hmgA gene of Pseudomonas aeruginosa leads to pyomelanin hyperproduction, stress resistance and increased persistence in chronic lung infection", Microbiology, 155, pp.1050~1057 (2009)
- Rosas ÁL, "Melanization affects susceptibility of Cryptococcus neoformans to heat and cold", FEMS Microbiol. Lett, 153, pp.265~272 (1997)
- Plonka PM, "Melanin synthesis in microorganisms - biotechnological and medical aspects", Acta Biochim. Pol, 53, pp.429~443 (2006)
- Turick CE, "Properties and function of pyomelanin", pp.449~472 (2010), Biopolymers
- Aronson JN, "Effects of m-tyrosine on growth and sporulation of Bacillus species", J. Bacteriol, 90, pp.38~46 (1965)
- Piñero S, "Tyrosinase from Rhizobium etli is involved in nodulation efficiency and symbiosis-associated stress resistance", J. Mol. Microbiol. Biotechnol, 13, pp.35~44 (2007)
- Solano F, "Isolation and characterization of strain MMB-1 (CECT 4803), a novel melanogenic marine bacterium", Appl. Environ. Microbiol, 63, pp.3499~3506 (1997)
- Coon SL, "Homogentisic acid is the product of MelA, which mediates melanogenesis in the marine bacterium Shewanella colwelliana D", Appl. Environ. Microbiol, 60, pp.3006~3010 (1994)
- Goodwin PH, "Brown pigmentation of Xanthomonas campestris pv. phaseoli associated with homogentisic acid", Can. J. Microbiol, 40, pp.28~34 (1994)
- Keith KE, "Burkholderia cenocepacia C5424 produces a pigment with antioxidant properties using a homogentisate intermediate", J. Bacteriol, 189, pp.9057~9065 (2007)
- Valeru SP, "Role of melanin pigment in expression of Vibrio cholerae virulence factors", Infect. Immun, 77, pp.935~942 (2009)
- Yabuuchi E, "Characterization of 'pyomelanin'-producing strains of Pseudomonas aeruginosa", Int. J. Syst. Evol. Microbiol, 22, pp.53~64 (1972)
- Hayward AC., "Encyclopedia of Microbiology", pp.32~42 (2000), Academic Press
- Salanoubat M, "Genome sequence of the plant pathogen Ralstonia solanacearum", Nature, 415, pp.497~502 (2002)
- Hernández-Romero D, "Polyphenol oxidase activity expression in Ralstonia solanacearum", Appl. Environ. Microbiol, 71, pp.6808~6815 (2005)
- Ahmad S, "Genetic determinants for pyomelanin production and its protective effect against oxidative stress in Ralstonia solanacearum", PLoS One, 11, pp.e0160845~ (2016)
- Jeong Y, "Genetic diversity and distribution of Korean isolates of Ralstonia solanacearum", Plant Dis, 91, pp.1277~1287 (2007)
- Hayward AC, "Laboratory Guide for Identification of Plant Pathogenic Bacteria", pp.151~173 (2001), APS Press
- Kelman A, "The relationship of pathogenicity of Pseudomonas solanacearum to colony appearance in a tetrazolium medium", Phytopathology, 44, pp.693~695 (1954)
- Wu J, "Loss of glutamate dehydrogenase in Ralstonia solanacearum alters dehydrogenase activity, extracellular polysaccharide production and bacterial virulence", Physiol. Mol. Plant Pathol, 90, pp.57~64 (2015)
- Yanisch-Perron C, "Improved M13phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors", Gene, 33, pp.103~119 (1985)
- Sambrook J, "Molecular Cloning:A Laboratory Manual" (1989), Cold Spring Harbor Laboratory
- Keen NT, "Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria", Gene, 70, pp.191~197 (1988)
- Boyer HW, "A complementation analysis of the restriction and modification of DNA in Escherichia coli", J. Mol. Biol, 41, pp.459~472 (1969)
- Figurski DH, "Replication of an origincontaining derivative of plasmid RK2 dependent on a plasmid function provided in trans", Proc. Natl. Acad. Sci. USA, 76, pp.1648~1652 (1979)
- Ngo HB, "The mitochondrial transcription and packaging factor Tfam imposes a U-turn on mitochondrial DNA", Nat. Struct. Mol. Biol, 18, pp.1290~1296 (2011)
- Denoya CD, "A Streptomyces avermitilis gene encoding a 4-hydroxyphenylpyruvic acid dioxygenase-like protein that directs the production of homogentisic acid and an ochronotic pigment in Escherichia coli", J. Bacteriol, 176, pp.5312~5319 (1994)
- Hunter RC, "A putative ABC transporter, hatABCDE, is among molecular determinants of pyomelanin production in Pseudomonas aeruginosa", J. Bacteriol, 192, pp.5962~5671 (2010)
- Ekstrand MI, "Mitochondrial transcription factor A regulates mtDNA copy number in mammals", Hum. Mol. Genet, 13, pp.935~944 (2004)
- Schmaler-Ripcke J, "Production of pyomelanin, a second type of melanin, via the tyrosine degradation pathway in Aspergillus fumigatus", Appl. Environ. Microbiol, 75, pp.493~503 (2009)
- Wan X, "Molecular and biochemical characterization of a distinct tyrosinase involved in melanin production from Aeromonas media", Appl. Microbiol. Biotechnol, 82, pp.261~269 (2009)
- Wang H, "Identification and molecular characterization of the homogentisate pathway responsible for pyomelanin production, the major melanin constituents in Aeromonas media WS", PLoS One, 10, pp.e0120923~ (2015)