caa a22 4500 445318090 CHVBK 20180317142654.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s10126-010-9269-z doi (NATIONALLICENCE)springer-10.1007/s10126-010-9269-z Molecular Cloning of Cecropin B Responsive Endonucleases in Yersinia ruckeri [Elektronische Daten] [Ulysses Sallum, Thomas Chen] We have previously demonstrated that Yersinia ruckeri resists cecropin B in an inducible manner. In this study, we sought to identify the molecular changes responsible for the inducible cecropin B resistance of Y. ruckeri. Differences in gene expression associated with the inducible resistance were investigated. Cultures of Y. ruckeri were exposed to a sublethal concentration of cecropin B and resultant changes in the messenger RNA population of the bacteria were assayed using the differential display reverse transcription polymerase chain reaction (DD-RT-PCR). A single band was consistently increased in intensity in all repeats of the experiment. The band was excised, cloned, sequenced, and used to screen a Y. ruckeri genomic DNA library. The DD-RT-PCR fragment shared 100% identity to the cDNA sequence of an ATP-dependent endonuclease of the overcome lysogenization defect (OLD) family of Y. ruckeri 29473. The genomic clone that was recovered was not identical to the DD-RT-PCR clone, but harbored a gene for a secreted endonuclease 1 (nucM) homologue. It was determined that transcription of the gene was upregulated following exposure to cecropin B via RT-PCR. Furthermore, an increase in the nuclease activity of culture supernatants of Y. ruckeri following exposure to cecropin B was demonstrated. These findings demonstrate that cecropin B exposure increases the expression of at least two endonucleases in Y. ruckeri. The production and secretion of an endonuclease by Y. ruckeri in response to an antimicrobial peptide indicates the involvement of both intracellular and extracellular DNA in the toxic effects of cecropin B. Springer Science+Business Media, LLC, 2010 Cecropin B nationallicence Yersinia ruckeri nationallicence Extracellular endonuclease nationallicence Antimicrobial peptide resistance nationallicence Neutrophil extracellular traps nationallicence Endonuclease 1 nationallicence ATP dependent endonuclease of the OLD family nationallicence Sallum Ulysses Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Rd., U-3125, 06269-3125, Storrs, CT, USA aut Chen Thomas Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Rd., U-3125, 06269-3125, Storrs, CT, USA aut Marine Biotechnology Springer-Verlag 13/1(2011-02-01), 56-65 1436-2228 13:1<56 2011 13 10126 https://doi.org/10.1007/s10126-010-9269-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9269-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sallum Ulysses Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Rd., U-3125, 06269-3125, Storrs, CT, USA aut NATIONALLICENCE 700 1- Chen Thomas Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Rd., U-3125, 06269-3125, Storrs, CT, USA aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/1(2011-02-01), 56-65 1436-2228 13:1<56 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer