Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery

Trans-kingdom conjugation (TKC)/inter-domain conjugation is a horizontal gene transfer phenomenon that transfers DNA from eubacteria to eukaryotes and archaebacteria via a type IV secretion system encoded in IncP1-type broad-host-range plasmids. Although TKC is considered a potential gene introducti...

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Main Authors: Kazuki Moriguchi, Kazuyuki Nakamura, Yudai Takahashi, Kyoko Higo-Moriguchi, Kazuya Kiyokawa, Katsunori Suzuki
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/3/488
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author Kazuki Moriguchi
Kazuyuki Nakamura
Yudai Takahashi
Kyoko Higo-Moriguchi
Kazuya Kiyokawa
Katsunori Suzuki
author_facet Kazuki Moriguchi
Kazuyuki Nakamura
Yudai Takahashi
Kyoko Higo-Moriguchi
Kazuya Kiyokawa
Katsunori Suzuki
author_sort Kazuki Moriguchi
collection DOAJ
description Trans-kingdom conjugation (TKC)/inter-domain conjugation is a horizontal gene transfer phenomenon that transfers DNA from eubacteria to eukaryotes and archaebacteria via a type IV secretion system encoded in IncP1-type broad-host-range plasmids. Although TKC is considered a potential gene introduction tool, donor chromosomal genes that influence TKC efficiency have rarely been analyzed, hindering targeted donor breeding. To identify potential TKC-related genes on a donor chromosome, a genome-wide screening of TKC-deficient mutants was performed using a comprehensive collection of <i>Escherichia coli</i> gene knockout mutants (Keio collection) as donors and a <i>Saccharomyces cerevisiae</i> strain as a recipient. Out of 3884 mutants, two mutants (∆<i>aceE</i>, ∆<i>priA</i>) showed a severe decrease in TKC efficiency by more than two orders of magnitude but not in bacterial conjugation. The effect on TKC efficiency by the two mutants was partly recovered by a preculture with a fresh culture medium before the TKC reaction, regardless of the presence of antibiotics. These results suggest that no single chromosomal target gene is solely responsible for universally blocking IncP1-type conjugation by impeding its function. The results also suggest the existence of an unidentified recognition or transfer mechanism distinct from bacterial conjugation, highlighting the novel roles of <i>aceE</i> and <i>priA</i>.
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spelling doaj-art-466fba983c6d4593b973eb0bee27f0782025-08-20T01:48:44ZengMDPI AGMicroorganisms2076-26072025-02-0113348810.3390/microorganisms13030488Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS MachineryKazuki Moriguchi0Kazuyuki Nakamura1Yudai Takahashi2Kyoko Higo-Moriguchi3Kazuya Kiyokawa4Katsunori Suzuki5Program of Basic Biology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima 739-8526, JapanProgram of Basic Biology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima 739-8526, JapanDepartment of Biological Science, Faculty of Science, Hiroshima University, Higashi-Hiroshima 739-8526, JapanFujita Health University School of Medicine, Toyoake 470-1192, JapanProgram of Basic Biology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima 739-8526, JapanProgram of Basic Biology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima 739-8526, JapanTrans-kingdom conjugation (TKC)/inter-domain conjugation is a horizontal gene transfer phenomenon that transfers DNA from eubacteria to eukaryotes and archaebacteria via a type IV secretion system encoded in IncP1-type broad-host-range plasmids. Although TKC is considered a potential gene introduction tool, donor chromosomal genes that influence TKC efficiency have rarely been analyzed, hindering targeted donor breeding. To identify potential TKC-related genes on a donor chromosome, a genome-wide screening of TKC-deficient mutants was performed using a comprehensive collection of <i>Escherichia coli</i> gene knockout mutants (Keio collection) as donors and a <i>Saccharomyces cerevisiae</i> strain as a recipient. Out of 3884 mutants, two mutants (∆<i>aceE</i>, ∆<i>priA</i>) showed a severe decrease in TKC efficiency by more than two orders of magnitude but not in bacterial conjugation. The effect on TKC efficiency by the two mutants was partly recovered by a preculture with a fresh culture medium before the TKC reaction, regardless of the presence of antibiotics. These results suggest that no single chromosomal target gene is solely responsible for universally blocking IncP1-type conjugation by impeding its function. The results also suggest the existence of an unidentified recognition or transfer mechanism distinct from bacterial conjugation, highlighting the novel roles of <i>aceE</i> and <i>priA</i>.https://www.mdpi.com/2076-2607/13/3/488genome-wide screeningIncP1-type plasmidtrans-kingdom conjugationtype IV secretion systemhorizontal gene transfer
spellingShingle Kazuki Moriguchi
Kazuyuki Nakamura
Yudai Takahashi
Kyoko Higo-Moriguchi
Kazuya Kiyokawa
Katsunori Suzuki
Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery
Microorganisms
genome-wide screening
IncP1-type plasmid
trans-kingdom conjugation
type IV secretion system
horizontal gene transfer
title Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery
title_full Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery
title_fullStr Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery
title_full_unstemmed Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery
title_short Genome-Wide Survey of Donor Chromosomal Genes Involved in Trans-Kingdom Conjugation via the RP4-T4SS Machinery
title_sort genome wide survey of donor chromosomal genes involved in trans kingdom conjugation via the rp4 t4ss machinery
topic genome-wide screening
IncP1-type plasmid
trans-kingdom conjugation
type IV secretion system
horizontal gene transfer
url https://www.mdpi.com/2076-2607/13/3/488
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