Detection of preQ0 deazaguanine modifications in bacteriophage CAjan DNA using Nanopore sequencing reveals same hypermodification at two distinct DNA motifs
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Detection of preQ0 deazaguanine modifications in bacteriophage CAjan DNA using Nanopore sequencing reveals same hypermodification at two distinct DNA motifs. / Kot, Witold; Olsen, Nikoline S.; Nielsen, Tue K.; Hutinet, Geoffrey; de Crecy-Lagard, Valerie; Cui, Liang; Dedon, Peter C.; Carstens, Alexander B.; Moineau, Sylvain; Swairjo, Manal A.; Hansen, Lars H.
In: Nucleic Acids Symposium Series, Vol. 48, No. 18, 2020, p. 10383-10396.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Detection of preQ0 deazaguanine modifications in bacteriophage CAjan DNA using Nanopore sequencing reveals same hypermodification at two distinct DNA motifs
AU - Kot, Witold
AU - Olsen, Nikoline S.
AU - Nielsen, Tue K.
AU - Hutinet, Geoffrey
AU - de Crecy-Lagard, Valerie
AU - Cui, Liang
AU - Dedon, Peter C.
AU - Carstens, Alexander B.
AU - Moineau, Sylvain
AU - Swairjo, Manal A.
AU - Hansen, Lars H.
PY - 2020
Y1 - 2020
N2 - In the constant evolutionary battle against mobile genetic elements (MGEs), bacteria have developed several defense mechanisms, some of which target the incoming, foreign nucleic acids e.g. restriction-modification (R-M) or CRISPR-Cas systems. Some of these MGEs, including bacteriophages, have in turn evolved different strategies to evade these hurdles. It was recently shown that the siphophage CAjan and 180 other viruses use 7-deazaguanine modifications in their DNA to evade bacterial R-M systems. Among others, phage CAjan genome contains a gene coding for a DNA-modifying homolog of a tRNA-deazapurine modification enzyme, together with four 7-cyano-7-deazaguanine synthesis genes. Using the CRISPR-Cas9 genome editing tool combined with the Nanopore Sequencing (ONT) we showed that the 7-deazaguanine modification in the CAjan genome is dependent on phage-encoded genes. The modification is also site-specific and is found mainly in two separate DNA sequence contexts: GA and GGC. Homology modeling of the modifying enzyme DpdA provides insight into its probable DNA binding surface and general mode of DNA recognition.
AB - In the constant evolutionary battle against mobile genetic elements (MGEs), bacteria have developed several defense mechanisms, some of which target the incoming, foreign nucleic acids e.g. restriction-modification (R-M) or CRISPR-Cas systems. Some of these MGEs, including bacteriophages, have in turn evolved different strategies to evade these hurdles. It was recently shown that the siphophage CAjan and 180 other viruses use 7-deazaguanine modifications in their DNA to evade bacterial R-M systems. Among others, phage CAjan genome contains a gene coding for a DNA-modifying homolog of a tRNA-deazapurine modification enzyme, together with four 7-cyano-7-deazaguanine synthesis genes. Using the CRISPR-Cas9 genome editing tool combined with the Nanopore Sequencing (ONT) we showed that the 7-deazaguanine modification in the CAjan genome is dependent on phage-encoded genes. The modification is also site-specific and is found mainly in two separate DNA sequence contexts: GA and GGC. Homology modeling of the modifying enzyme DpdA provides insight into its probable DNA binding surface and general mode of DNA recognition.
KW - CRYSTAL-STRUCTURE
KW - REPAIR
KW - BASE
U2 - 10.1093/nar/gkaa735
DO - 10.1093/nar/gkaa735
M3 - Journal article
C2 - 32941607
VL - 48
SP - 10383
EP - 10396
JO - Nucleic acids symposium series
JF - Nucleic acids symposium series
SN - 0261-3166
IS - 18
ER -
ID: 253732970