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      Excisionase in Pf filamentous prophage controls lysis‐lysogeny decision‐making in Pseudomonas aeruginosa

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          Summary

          Pf filamentous prophages are prevalent among clinical and environmental Pseudomonas aeruginosa isolates. Pf4 and Pf5 prophages are integrated into the host genomes of PAO1 and PA14, respectively, and play an important role in biofilm development. However, the genetic factors that directly control the lysis‐lysogeny switch in Pf prophages remain unclear. Here, we identified and characterized the excisionase genes in Pf4 and Pf5 (named xisF4 and xisF5, respectively). XisF4 and XisF5 represent two major subfamilies of functional excisionases and are commonly found in Pf prophages. While both of them can significantly promote prophage excision, only XisF5 is essential for Pf5 excision. XisF4 activates Pf4 phage replication by upregulating the phage initiator gene ( PA0727). In addition, xisF4 and the neighboring phage repressor c gene pf4r are transcribed divergently and their 5′‐untranslated regions overlap. XisF4 and Pf4r not only auto‐activate their own expression but also repress each other. Furthermore, two H‐NS family proteins, MvaT and MvaU, coordinately repress Pf4 production by directly repressing xisF4. Collectively, we reveal that Pf prophage excisionases cooperate in controlling lysogeny and phage production.

          Abstract

          A proposed model of the lysis‐lysogeny switch of Pf4 in PAO1. Integration of Pf4 in host genome is maintained by MvaT/MvaU by binding to the excisionase gene xisF4. Inactivation of MvaT/MvaU upregulates xisF4 and subsequently, XisF4 activates the replication initiator gene PA0727, leading to Pf4 phage replication and release.

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          Most cited references43

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          An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants.

          Random transposon insertion libraries have proven invaluable in studying bacterial genomes. Libraries that approach saturation must be large, with multiple insertions per gene, making comprehensive genome-wide scanning difficult. To facilitate genome-scale study of the opportunistic human pathogen Pseudomonas aeruginosa strain PA14, we constructed a nonredundant library of PA14 transposon mutants (the PA14NR Set) in which nonessential PA14 genes are represented by a single transposon insertion chosen from a comprehensive library of insertion mutants. The parental library of PA14 transposon insertion mutants was generated by using MAR2xT7, a transposon compatible with transposon-site hybridization and based on mariner. The transposon-site hybridization genetic footprinting feature broadens the utility of the library by allowing pooled MAR2xT7 mutants to be individually tracked under different experimental conditions. A public, internet-accessible database (the PA14 Transposon Insertion Mutant Database, http://ausubellab.mgh.harvard.edu/cgi-bin/pa14/home.cgi) was developed to facilitate construction, distribution, and use of the PA14NR Set. The usefulness of the PA14NR Set in genome-wide scanning for phenotypic mutants was validated in a screen for attachment to abiotic surfaces. Comparison of the genes disrupted in the PA14 transposon insertion library with an independently constructed insertion library in P. aeruginosa strain PAO1 provides an estimate of the number of P. aeruginosa essential genes.
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            The SOS regulatory system of Escherichia coli.

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              PBAD-based shuttle vectors for functional analysis of toxic and highly regulated genes in Pseudomonas and Burkholderia spp. and other bacteria.

              We report the construction of a series of Escherichia-Pseudomonas broad-host-range expression vectors utilizing the P(BAD) promoter and the araC regulator for routine cloning, conditional expression, and analysis of tightly controlled and/or toxic genes in pseudomonads.
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                Author and article information

                Contributors
                xxwang@scsio.ac.cn
                Journal
                Mol Microbiol
                Mol. Microbiol
                10.1111/(ISSN)1365-2958
                MMI
                Molecular Microbiology
                John Wiley and Sons Inc. (Hoboken )
                0950-382X
                1365-2958
                12 December 2018
                February 2019
                : 111
                : 2 ( doiID: 10.1111/mmi.2019.111.issue-2 )
                : 495-513
                Affiliations
                [ 1 ] Key Laboratory of Tropical Marine Bio‐resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology Chinese Academy of Sciences Guangzhou 510301 PR China
                [ 2 ] University of Chinese Academy of Sciences Beijing 100049 China
                Author notes
                [*] [* ] For correspondence. E‐mail xxwang@ 123456scsio.ac.cn ; Tel. +86 20 89267515; Fax +86 20 89235490.

                Author information
                https://orcid.org/0000-0002-2006-1746
                Article
                MMI14170
                10.1111/mmi.14170
                7379572
                30475408
                2444c5e3-d686-4df1-a361-fa672dbc83b1
                © 2018 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 17 November 2018
                Page count
                Figures: 9, Tables: 1, Pages: 19, Words: 25961
                Funding
                Funded by: National Natural Science Foundation of China , open-funder-registry 10.13039/501100001809;
                Award ID: 31625001
                Award ID: 31500025
                Award ID: 41706172
                Funded by: National Basic Research Program of China
                Award ID: 2017YFC0506303
                Award ID: 2018YFC1406500
                Funded by: Strategic Priority Research Program of the Chinese Academy of Sciences
                Award ID: XDA13010500
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                February 2019
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.5 mode:remove_FC converted:24.07.2020

                Microbiology & Virology
                Microbiology & Virology

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