A novel resistance gene, lnu(H), conferring resistance to lincosamides in Riemerella anatipestifer CH-2

•First report of lnu(H), a novel lincosamide nucleotidylyltransferase gene.•lnu(H) confers high-level lincosamide resistance in Riemerella anatipestifer CH-2.•Lnu(H) catalysed adenylylation of lincosamides as demonstrated by mass spectrometry.•lnu(H) is widely distributed and transferrable among R....

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Published in:International journal of antimicrobial agents 2018-01, Vol.51 (1), p.136-139
Main Authors: Luo, Hong-Yan, Liu, Ma-Feng, Wang, Ming-Shu, Zhao, Xin-Xin, Jia, Ren-Yong, Chen, Shun, Sun, Kun-Feng, Yang, Qiao, Wu, Ying, Chen, Xiao-Yue, Biville, Francis, Zou, Yuan-Feng, Jing, Bo, Cheng, An-Chun, Zhu, De-Kang
Format: Article
Language:English
Subjects:
Lincosamide resistance
lnu(H)
Nucleotidylyltransferase
Riemerella anatipestifer
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Summary:•First report of lnu(H), a novel lincosamide nucleotidylyltransferase gene.•lnu(H) confers high-level lincosamide resistance in Riemerella anatipestifer CH-2.•Lnu(H) catalysed adenylylation of lincosamides as demonstrated by mass spectrometry.•lnu(H) is widely distributed and transferrable among R. anatipestifer isolates. The Gram-negative bacterium Riemerella anatipestifer CH-2 is resistant to lincosamides, having a lincomycin (LCM) minimum inhibitory concentration (MIC) of 128 µg/mL. The G148_1775 gene of R. anatipestifer CH-2, designated lnu(H), encodes a 260-amino acid protein with ≤41% identity to other reported lincosamide nucleotidylyltransferases. Escherichia coli RosettaTM (DE3) containing the pBAD24-lnu(H) plasmid showed four- and two-fold increases in the MICs of LCM and clindamycin (CLI), respectively. A kinetic assay of the purified Lnu(H) enzyme for LCM and CLI showed that the protein could inactive lincosamides. Mass spectrometry analysis demonstrated that the Lnu(H) enzyme catalysed adenylylation of lincosamides. In addition, an lnu(H) gene deletion strain exhibited 512- and 32-fold decreases in LCM and CLI MICs, respectively. The wild-type level of lincosamide resistance could be restored by complementation with a shuttle plasmid carrying the lnu(H) gene. The transformant R. anatipestifer ATCC 11845 [lnu(H)] acquired by natural transformation also exhibited high-level lincosamide resistance. Moreover, among 175 R. anatipestifer field isolates, 56 (32.0%) were positive for the lnu(H) gene by PCR. In conclusion, Lnu(H) is a novel lincosamide nucleotidylyltransferase that inactivates LCM and CLI by nucleotidylylation, thus conferring high-level lincosamide resistance to R. anatipestifer CH-2.
ISSN:0924-8579
1872-7913
DOI:10.1016/j.ijantimicag.2017.08.022