Two Distinct Modes of Lysis Regulation in Campylobacter Fletchervirus and Firehammervirus Phages
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Two Distinct Modes of Lysis Regulation in Campylobacter Fletchervirus and Firehammervirus Phages. / Zampara, Athina; Ahern, Stephen J.; Briers, Yves; Brøndsted, Lone; Sørensen, Martine Camilla Holst.
I: Viruses, Bind 12, Nr. 11, 1247, 2020.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Two Distinct Modes of Lysis Regulation in Campylobacter Fletchervirus and Firehammervirus Phages
AU - Zampara, Athina
AU - Ahern, Stephen J.
AU - Briers, Yves
AU - Brøndsted, Lone
AU - Sørensen, Martine Camilla Holst
PY - 2020
Y1 - 2020
N2 - Campylobacter phages are divided into two genera; Fletchervirus and Firehammervirus, showing only limited intergenus homology. Here, we aim to identify the lytic genes of both genera using two representative phages (F352 and F379) from our collection. We performed a detailed in silico analysis searching for conserved protein domains and found that the predicted lytic genes are not organized into lysis cassettes but are conserved within each genus. To verify the function of selected lytic genes, the proteins were expressed in E. coli, followed by lytic assays. Our results show that Fletchervirus phages encode a typical signal peptide (SP) endolysin dependent on the Sec-pathway for translocation and a holin for activation. In contrast, Firehammervirus phages encode a novel endolysin that does not belong to currently described endolysin groups. This endolysin also uses the Sec-pathway for translocation but induces lysis of E. coli after overexpression. Interestingly, co-expression of this endolysin with an overlapping gene delayed and limited cell lysis, suggesting that this gene functions as a lysis inhibitor. These results indicate that Firehammervirus phages regulate lysis timing by a yet undescribed mechanism. In conclusion, we found that the two Campylobacter phage genera control lysis by two distinct mechanisms.
AB - Campylobacter phages are divided into two genera; Fletchervirus and Firehammervirus, showing only limited intergenus homology. Here, we aim to identify the lytic genes of both genera using two representative phages (F352 and F379) from our collection. We performed a detailed in silico analysis searching for conserved protein domains and found that the predicted lytic genes are not organized into lysis cassettes but are conserved within each genus. To verify the function of selected lytic genes, the proteins were expressed in E. coli, followed by lytic assays. Our results show that Fletchervirus phages encode a typical signal peptide (SP) endolysin dependent on the Sec-pathway for translocation and a holin for activation. In contrast, Firehammervirus phages encode a novel endolysin that does not belong to currently described endolysin groups. This endolysin also uses the Sec-pathway for translocation but induces lysis of E. coli after overexpression. Interestingly, co-expression of this endolysin with an overlapping gene delayed and limited cell lysis, suggesting that this gene functions as a lysis inhibitor. These results indicate that Firehammervirus phages regulate lysis timing by a yet undescribed mechanism. In conclusion, we found that the two Campylobacter phage genera control lysis by two distinct mechanisms.
KW - Campylobacter phages
KW - endolysins
KW - lysis inhibitor
KW - lysis regulation
U2 - 10.3390/v12111247
DO - 10.3390/v12111247
M3 - Journal article
C2 - 33142851
AN - SCOPUS:85095570959
VL - 12
JO - Viruses
JF - Viruses
SN - 1999-4915
IS - 11
M1 - 1247
ER -
ID: 251586064