Genome-Wide Transcriptional Start Site Mapping and sRNA Identification in the PathogenLeptospira interrogans.
Leptospiraare emerging zoonotic pathogens transmitted from animals to humans typically through contaminated environmental sources of water and soil. Regulatory pathways of pathogenicLeptospiraspp. underlying the adaptive response to different hosts and environmental conditions remains elusive. In this study, we provide the first global Transcriptional Start Site (TSS) map of aLeptospiraspecies. RNA was obtained from the pathogenLeptospira interrogansgrown at 30°C (optimalin vitrotemperature) and 37°C (host temperature) and selectively enriched for 5' ends of native transcripts. A total of 2865 and 2866 primary TSS (pTSS) were predicted in the genome ofL. interrogansat 30 and 37°C, respectively. The majority of the pTSSs were located between 0 and 10 nucleotides from the translational start site, suggesting that leaderless transcripts are a common feature of the leptospiral translational landscape. Comparative differential RNA-sequencing (dRNA-seq) analysis revealed conservation of most pTSS at 30 and 37°C. Promoter prediction algorithms allow the identification of the binding sites of the alternative sigma factor sigma 54. However, other motifs were not identified indicating thatLeptospiraconsensus promoter sequences are inherently different from theEscherichia colimodel. RNA sequencing also identified 277 and 226 putative small regulatory RNAs (sRNAs) at 30 and 37°C, respectively, including eight validated sRNAs by Northern blots. These results provide the first global view of TSS and the repertoire of sRNAs inL. interrogans. These data will establish a foundation for future experimental work on gene regulation under various environmental conditions including those in the host.
Bioinformatics and Biostatistics Hub, Institut Pasteur, C3BI Paris, France.
Anna
A
Zhukova
Biology of Spirochetes Unit, Institut Pasteur Paris, France.
Luis Guilherme
LG
Fernandes
Biology of Spirochetes Unit, Institut PasteurParis, France; Mutualized Microbiology Platform, Institut Pasteur, Pasteur International Bioresources NetworkParis, France.
Perrine
P
Hugon
Biology of Spirochetes Unit, Institut PasteurParis, France; Department of Biology, Manhattanville CollegePurchase, NY, USA.
Christopher J
CJ
Pappas
CITECH, Institut Pasteur, Plate-forme Transcriptome et Epigenome, Pole Biomics - CITECH Paris, France.
Odile
O
Sismeiro
CITECH, Institut Pasteur, Plate-forme Transcriptome et Epigenome, Pole Biomics - CITECH Paris, France.
Jean-Yves
JY
Coppée
Bioinformatics and Biostatistics Hub, Institut Pasteur, C3BI Paris, France.
Christophe
C
Becavin
Bioinformatics and Biostatistics Hub, Institut Pasteur, C3BI Paris, France.
Christophe
C
Malabat
Biology of Spirochetes Unit, Institut Pasteur Paris, France.
Azad
A
Eshghi
Department of Microbiology and Immunology, Indiana University School of Medicine Indianapolis, IN, USA.
Jun-Jie
JJ
Zhang
Department of Microbiology and Immunology, Indiana University School of Medicine Indianapolis, IN, USA.
Frank X
FX
Yang
Biology of Spirochetes Unit, Institut Pasteur Paris, France.
Mathieu
M
Picardeau
D002874:Chromosome Mapping; D016680:Genome, Bacterial; D007921:Leptospira interrogans / Q000235:genetics* / Q000254:growth & development; D058727:RNA, Small Untranslated / Q000235:genetics*; D013696:Temperature; D024363:Transcription Initiation Site
D058727:RNA, Small Untranslated
RNA; leptospirosis; promoter; spirochetes; transcription factors