TY  - JOUR
A1  - Westermann, Alexander J.
A1  - Barquist, Lars
A1  - Vogel, Jörg
T1  - Resolving host-pathogen interactions by dual RNA-seq
JF  - PLoS Pathogens
N2  - The transcriptome is a powerful proxy for the physiological state of a cell, healthy or diseased. As a result, transcriptome analysis has become a key tool in understanding the molecular changes that accompany bacterial infections of eukaryotic cells. Until recently, such transcriptomic studies have been technically limited to analyzing mRNA expression changes in either the bacterial pathogen or the infected eukaryotic host cell. However, the increasing sensitivity of high-throughput RNA sequencing now enables “dual RNA-seq” studies, simultaneously capturing all classes of coding and noncoding transcripts in both the pathogen and the host. In the five years since the concept of dual RNA-seq was introduced, the technique has been applied to a range of infection models. This has not only led to a better understanding of the physiological changes in pathogen and host during the course of an infection but has also revealed hidden molecular phenotypes of virulence-associated small noncoding RNAs that were not visible in standard infection assays. Here, we use the knowledge gained from these recent studies to suggest experimental and computational guidelines for the design of future dual RNA-seq studies. We conclude this review by discussing prospective applications of the technique.
KW  - Medicine
KW  - RNA sequencing
KW  - Salmonellosis
KW  - Transcriptome analysis
KW  - Gene expression
KW  - Bacterial pathogens
KW  - Salmonella
KW  - Host cells
KW  - Lysis (medicine)
Y1  - 2017
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-171921
VL  - 13
IS  - 2
ER  - 
TY  - JOUR
A1  - Lioliou, Efthimia
A1  - Sharma, Cynthia M.
A1  - Caldelari, Isabelle
A1  - Helfer, Anne-Catherine
A1  - Fechter, Pierre
A1  - Vandenesch, François
A1  - Vogel, Jörg
A1  - Romby, Pascale
T1  - Global Regulatory Functions of the Staphylococcus aureus Endoribonuclease III in Gene Expression
JF  - PLoS Genetics
N2  - RNA turnover plays an important role in both virulence and adaptation to stress in the Gram-positive human pathogen Staphylococcus aureus. However, the molecular players and mechanisms involved in these processes are poorly understood. Here, we explored the functions of S. aureus endoribonuclease III (RNase III), a member of the ubiquitous family of double-strand-specific endoribonucleases. To define genomic transcripts that are bound and processed by RNase III, we performed deep sequencing on cDNA libraries generated from RNAs that were co-immunoprecipitated with wild-type RNase III or two different cleavage-defective mutant variants in vivo. Several newly identified RNase III targets were validated by independent experimental methods. We identified various classes of structured RNAs as RNase III substrates and demonstrated that this enzyme is involved in the maturation of rRNAs and tRNAs, regulates the turnover of mRNAs and non-coding RNAs, and autoregulates its synthesis by cleaving within the coding region of its own mRNA. Moreover, we identified a positive effect of RNase III on protein synthesis based on novel mechanisms. RNase III–mediated cleavage in the 5′ untranslated region (5′UTR) enhanced the stability and translation of cspA mRNA, which encodes the major cold-shock protein. Furthermore, RNase III cleaved overlapping 5′UTRs of divergently transcribed genes to generate leaderless mRNAs, which constitutes a novel way to co-regulate neighboring genes. In agreement with recent findings, low abundance antisense RNAs covering 44% of the annotated genes were captured by co-immunoprecipitation with RNase III mutant proteins. Thus, in addition to gene regulation, RNase III is associated with RNA quality control of pervasive transcription. Overall, this study illustrates the complexity of post-transcriptional regulation mediated by RNase III.
KW  - staphylococcus aureus
KW  - ribonucleases
KW  - messenger RNA
KW  - RNA sequencing
KW  - antisense RNA
KW  - RNA structure
KW  - RNA synthesis
KW  - RNA denaturation
Y1  - 2012
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-127219
VL  - 8
IS  - 6
ER  -