Abstract
Unlike Ff-like coliphages, certain filamentous Inoviridae phages integrate their genomes into the host chromosome and enter a prophage state in their infectious cycle. This lysogenic life cycle was first reported for Xanthomonas citri Cf phage. However, except for the X. citri phages Cf and XacF1, complete genome sequence information about lysogenic Xanthomonas phages is not available to date. A proviral sequence of Xf109 phage was identified in the genome of Xanthomonas oryzae, the rice bacterial blight pathogen, and revived as infectious virions to lysogenize its host de novo. The genome of Xf109 phage is 7190 nucleotides in size and contains 12 predicted open reading frames in an organization similar to that of the Cf phage genome. Seven of the Xf109 proteins show significant sequence similarity to Cf and XacF1 phage proteins, while its ORF4 shares 92 % identity with the major coat protein of X. phage oryzae Xf. Integration of Xf109 phage DNA into the host genome is site-specific, and the attP/attB sequence contains the dif core sequence 5’-TATACATTATGCGAA-3’, which is identical to that of Cf, XacF1, and Xanthomonas campestris phage ϕLf. To my knowledge, this is the first complete genome sequence of a filamentous bacteriophage that infects X. oryzae.
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References
- 1.
Ahmad AA, Askora A, Kawasaki T, Fujie M, Yamada T (2014) The filamentous phage XacF1 causes loss of virulence in Xanthomonas axonopodis pv. citri, the causative agent of citrus canker disease. Front Microbiol 5:321
- 2.
Campos J, Martinez E, Suzarte E, Rodriguez BL, Marrero K, Silva Y, Ledon T, del Sol R, Fando R (2003) VGJϕ, a novel filamentous phage of Vibrio cholerae, integrates into the same chromosomal site as CTXϕ. J Bacteriol 185:5685–5696
- 3.
Dai H, Tsay SH, Kuo TT, Lin YH, Wu WC (1987) Neolysogenization of Xanthomonas campestris pv. citri infected with filamentous phage Cf16. Virology 156:313–320
- 4.
Dai H, Chow TY, Liao HJ, Chen ZY, Chiang KS (1988) Nucleotide sequences involved in the neolysogenic insertion of filamentous phage Cf16-v1 into the Xanthomonas campestris pv. citri chromosome. Virology 167:613–620
- 5.
Das B, Bischerour J, Val ME, Barre FX (2010) Molecular keys of the tropism of integration of the cholera toxin phage. Proc Natl Acad Sci USA 107:4377–4382
- 6.
Das B, Bischerour J, Barre FX (2011) VGJϕ integration and excision mechanisms contribute to the genetic diversity of Vibrio cholerae epidemic strains. Proc Natl Acad Sci USA 108:2516–2521
- 7.
Frangione B, Nakashima Y, Konigsberg W, Wiseman RL (1978) The amino acid sequence of the major coat protein subunit of the filamentous virus Xf. FEBS Lett 96:381–384
- 8.
Fu JF, Chang RY, Tseng YH (1992) Construction of stable lactose-utilizing Xanthomonas campestris by chromosomal integration of cloned lac genes using filamentous phage ϕLf DNA. Appl Microbiol and Biotechnol 37:225–229
- 9.
Gonzalez MD, Lichtensteiger CA, Caughlan R, Vimr ER (2002) Conserved filamentous prophage in Escherichia coli O18:K1:H7 and Yersinia pestis biovar orientalis. J Bacteriol 184:6050–6055
- 10.
Heilpern AJ, Waldor MK (2003) pIIICTX, a predicted CTXϕ minor coat protein, can expand the host range of coliphage fd to include Vibrio cholerae. J Bacteriol 185:1037–1044
- 11.
Huber KE, Waldor MK (2002) Filamentous phage integration requires the host recombinases XerC and XerD. Nature 417:656–659
- 12.
Kamiunten H, Wakimoto S (1980) Effect of the infection with filamentous phage Xf2 on the properties of Xanthomonas campestris pv. oryzae. Jpn J Phytopathol 47:627–636
- 13.
Kamiunten H (1988) Restriction maps of filamentous phage Xf and Xf2 DNA. Jpn J Phytopathol 54:571–576
- 14.
Kamiunten H (1995) Integration of filamentous phage Xf2 DNA into chromosomal DNA of Xanthomonas campestris pv. oryzae. Jpn J Phytopathol 61:127–129
- 15.
Kawasaki T, Nagata S, Fujiwara A, Satsuma H, Fujie M, Usami S, Yamada T (2007) Genomic characterization of the filamentous integrative bacteriophages ϕRSS1 and ϕRSM1, which infect Ralstonia solanacearum. J Bacteriol 189:5792–5802
- 16.
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874
- 17.
Kuo TT, Huang TC, Wu RY, Yang CM (1967) Characterization of three bacteriophages of Xanthomonas oryzae (Uyeda Et Ishiyama) Dowson. Bot Bull Acad Sin 8:246–254
- 18.
Kuo TT, Huang TC, Chow TY (1969) A filamentous bacteriophage from Xanthomonas oryzae. Virology 39:548–555
- 19.
Kuo TT, Chao YS, Lin YH, Lin BY, Liu LF, Feng TY (1987) Integration of the DNA of filamentous bacteriophage Cflt into the chromosomal DNA of its host. J Virol 61:60–65
- 20.
Kuo TT, Lin YH, Huang CM, Chang SF, Dai H, Feng TY (1987) The lysogenic cycle of the filamentous phage Cflt from Xanthomonas campestris pv. citri. Virology 156:305–312
- 21.
Kuo TT, Tan MS, Su MT, Yang MK (1991) Complete nucleotide sequence of filamentous phage Cf1c from Xanthomonas campestris pv. citri. Nucleic Acids Res 19:2498
- 22.
Lee SJ (1991) Sequences that mediate the integration of ϕLf DNA into the chromosome of Xanthomonas campestris pv. campestris. National Chung Hsing University, National Chung Hsing University
- 23.
Lin NT, You BY, Huang CY, Kuo CW, Wen FS, Yang JS, Tseng YH (1994) Characterization of two novel filamentous phages of Xanthomonas. J Gen Virol 75:2543–2547
- 24.
Lin NT, Wen FS, Tseng YH (1996) A region of the filamentous phage ϕLf genome that can support autonomous replication and miniphage production. Biochem Biophys Res Commun 218:12–16
- 25.
Lin NT, Liu TJ, Lee TC, You BY, Yang MH, Wen FS, Tseng YH (1999) The adsorption protein genes of Xanthomonas campestris filamentous phages determining host specificity. J Bacteriol 181:2465–2471
- 26.
Liu TJ, Wen FS, Tseng TT, Yang MT, Lin NT, Tseng YH (1997) Identification of gene VI of filamentous phage ϕLf coding for a 10-kDa minor coat protein. Biochem Biophys Res Commun 239:752–755
- 27.
Luiten RG, Putterman DG, Schoenmakers JG, Konings RN, Day LA (1985) Nucleotide sequence of the genome of Pf3, an IncP-1 plasmid-specific filamentous bacteriophage of Pseudomonas aeruginosa. J Virol 56:268–276
- 28.
Marvin DA, Hohn B (1969) Filamentous bacterial viruses. Bacteriol Rev 33:172–209
- 29.
Nino-Liu DO, Ronald PC, Bogdanove AJ (2006) Xanthomonas oryzae pathovars: model pathogens of a model crop. Mol Plant Pathol 7:303–324
- 30.
Petrova M, Shcherbatova N, Kurakov A, Mindlin S (2014) Genomic characterization and integrative properties of ϕSMA6 and ϕSMA7, two novel filamentous bacteriophages of Stenotrophomonas maltophilia. Arch Virol 159:1293–1303
- 31.
Shieh GJ, Charng YC, Yang BC, Jenn T, Bau HJ, Kuo TT (1991) Identification and nucleotide sequence analysis of an open reading frame involved in high-frequency conversion of turbid to clear plaque mutants of filamentous phage Cf1t. Virology 185:316–322
- 32.
Val ME, Bouvier M, Campos J, Sherratt D, Cornet F, Mazel D, Barre FX (2005) The single-stranded genome of phage CTX is the form used for integration into the genome of Vibrio cholerae. Mol Cell 19:559–566
- 33.
Wen FS, Tseng YH (1994) Nucleotide sequence determination, characterization and purification of the single-stranded DNA-binding protein and major coat protein of filamentous phage ϕLf of Xanthomonas campestris pv. campestris. J Gen Virol 75:15–22
- 34.
Wen FS, Tseng YH (1996) Nucleotide sequence of the gene presumably encoding the adsorption protein of filamentous phage ϕLf. Gene 172:161–162
- 35.
Yang MK, Su WC, Kuo TT (1991) Highly efficient transfection of Xanthomonas campestris by electroporation. Bot Bull Acad Sin 32:197–203
- 36.
Yen MR, Lin NT, Hung CH, Choy KT, Weng SF, Tseng YH (2002) oriC region and replication termination site, dif, of the Xanthomonas campestris pv. campestris 17 chromosome. Appl Environ Microbiol 68:2924–2933
Acknowledgments
I thank Mr. Gregory Contreras for his help with data analysis and English editing.
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This study was supported by Career Development and Bridge Research Fund from Auxergen.
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Fig. S1
Treatment of Xf109 DNA with hydrolytic enzymes followed by electrophoresis in a 1 % agarose gel in Tris-acetate-EDTA buffer and staining in ethidium bromide with bovine pancreas RNase A (1.5 μg/ml) for 15 minutes. (A) Lane 1, DNA marker; lanes 2 and 3, DNA from Xf109 phage particles (200 ng) treated with S1 nuclease and DNase I, respectively; lane 4, untreated phage DNA. (B) Lane 1, DNA marker; lane 2, Xf109 RF DNA cut with EcoRV; lane 3, uncut Xf109 RF DNA (TIFF 2188 kb)
Fig. S2
The predicted signal peptide in Xf109 ORF4 protein. The output score (C-, S-, Y-, and D-score) format is described in the SignalP 4.1 server (http://www.cbs.dtu.dk/services/SignalP /output.php). The cleavage site of the signal peptide is predicted to be between alanine 30 (gray) and serine 31 (black) based on the D-score (D=0.826). The cutoff D-score is 0.510 (pink line) (TIFF 7067 kb)
Fig. S3
Phylogenic analysis of Xf109 ORF5, phage absorption protein pIII of ϕLf (Q37972, [34]), ϕXv (AF069776), and ϕXo (AF162859), Cf1cp3 of Cf1c and ORF6 of XacF1. (A) The percent identity matrix was computed using the Clustal Omega program. (B) Phylogenic tree inferred using the neighbor-joining method and constructed using MEGA7 software [16]. The optimal tree with a sum of branch lengths of 6.71675156 is shown. The evolutionary distances were computed using the Poisson correction method. All positions containing gaps and missing data were eliminated. There were a total of 328 positions in the final dataset (TIFF 4001 kb)
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Yeh, T.Y. Complete nucleotide sequence of a new filamentous phage, Xf109, which integrates its genome into the chromosomal DNA of Xanthomonas oryzae . Arch Virol 162, 567–572 (2017). https://doi.org/10.1007/s00705-016-3105-3
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Keywords
- Complete Genome Sequence
- Yersinia Pestis
- Filamentous Phage
- attP Site
- Replication Initiation Protein