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The novel cis-encoded antisense RNA AsrC positively regulates the expression of rpoE-rseABC operon and thus enhances the motility of Salmonella enterica serovar typhi.

Zhang Q, Zhang Y, Zhang X, Zhan L, Zhao X, Xu S, Sheng X, Huang X - Front Microbiol (2015)

Bottom Line: We found that AsrC increased the levels of rseC mRNA and protein.The expression of rpoE was also increased in S. typhi after overexpression of AsrC, which was dependent on rseC.Thus, we propose that AsrC increased RseC level and indirectly activating RpoE which can initiate fliA expression and promote the motility of S. typhi.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Jiangsu University - School of Medicine Zhenjiang, China ; Danyang People's Hospital of Jiangsu Province Danyang, China.

ABSTRACT
Bacterial non-coding RNAs are essential in many cellular processes, including response to environmental stress, and virulence. Deep sequencing analysis of the Salmonella enterica serovar typhi (S. typhi) transcriptome revealed a novel antisense RNA transcribed in cis on the strand complementary to rseC, an activator gene of sigma factor RpoE. In this study, expression of this antisense RNA was confirmed in S. typhi by Northern hybridization. Rapid amplification of cDNA ends and sequence analysis identified an 893 bp sequence from the antisense RNA coding region that covered all of the rseC coding region in the reverse direction of transcription. This sequence of RNA was named as AsrC. After overexpression of AsrC with recombinantant plasmid in S. typhi, the bacterial motility was increased obviously. To explore the mechanism of AsrC function, regulation of rseC and rpoE expression by AsrC was investigated. We found that AsrC increased the levels of rseC mRNA and protein. The expression of rpoE was also increased in S. typhi after overexpression of AsrC, which was dependent on rseC. Thus, we propose that AsrC increased RseC level and indirectly activating RpoE which can initiate fliA expression and promote the motility of S. typhi.

No MeSH data available.


Schematic representation and sequence analyses of asrC. (A) Genomic location of asrC. Black arrows, location of rseB, rseC, and lepA genes; gray arrows, position of asrC. (B) Sequence analysis by RACE of the AsrC. Bent arrow, transcription start site; +1 and ▲, experimentally determined transcription start site and transcription stop site of the asrC, respectively; underlined sequence, two different positions of oligonucleotide probes used for Northern hybridization for AsrC expression.
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Figure 1: Schematic representation and sequence analyses of asrC. (A) Genomic location of asrC. Black arrows, location of rseB, rseC, and lepA genes; gray arrows, position of asrC. (B) Sequence analysis by RACE of the AsrC. Bent arrow, transcription start site; +1 and ▲, experimentally determined transcription start site and transcription stop site of the asrC, respectively; underlined sequence, two different positions of oligonucleotide probes used for Northern hybridization for AsrC expression.

Mentions: By transcriptome analysis of S. typhi, several new ncRNAs were found, including a putative antisense RNA for rseC. To determine the boundaries of the putative novel ncRNA, rapid amplification of cDNA ends (RACEs) was used. Results of 5′-RACE and 3′-RACE revealed the 5′-end of the transcript, which was 119 bp downstream of the lepA start codon. The 3′-end of the transcript was 36 bp upstream of the rseC start codon, indicating a full-length transcript of 893 nt. The gene structure and sequence of the transcript are in Figure 1. End mapping revealed that the novel transcript was transcribed in cis from the strand complementary to rseC and overlapped the entire rseC mRNA. We therefore named it AsrC. From the entire sequence of the novel asRNA, no obvious ORF structure longer than 150 nt or SD element was found. Thus, we hypothesized that the novel asRNA was an ncRNA.


The novel cis-encoded antisense RNA AsrC positively regulates the expression of rpoE-rseABC operon and thus enhances the motility of Salmonella enterica serovar typhi.

Zhang Q, Zhang Y, Zhang X, Zhan L, Zhao X, Xu S, Sheng X, Huang X - Front Microbiol (2015)

Schematic representation and sequence analyses of asrC. (A) Genomic location of asrC. Black arrows, location of rseB, rseC, and lepA genes; gray arrows, position of asrC. (B) Sequence analysis by RACE of the AsrC. Bent arrow, transcription start site; +1 and ▲, experimentally determined transcription start site and transcription stop site of the asrC, respectively; underlined sequence, two different positions of oligonucleotide probes used for Northern hybridization for AsrC expression.
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Figure 1: Schematic representation and sequence analyses of asrC. (A) Genomic location of asrC. Black arrows, location of rseB, rseC, and lepA genes; gray arrows, position of asrC. (B) Sequence analysis by RACE of the AsrC. Bent arrow, transcription start site; +1 and ▲, experimentally determined transcription start site and transcription stop site of the asrC, respectively; underlined sequence, two different positions of oligonucleotide probes used for Northern hybridization for AsrC expression.
Mentions: By transcriptome analysis of S. typhi, several new ncRNAs were found, including a putative antisense RNA for rseC. To determine the boundaries of the putative novel ncRNA, rapid amplification of cDNA ends (RACEs) was used. Results of 5′-RACE and 3′-RACE revealed the 5′-end of the transcript, which was 119 bp downstream of the lepA start codon. The 3′-end of the transcript was 36 bp upstream of the rseC start codon, indicating a full-length transcript of 893 nt. The gene structure and sequence of the transcript are in Figure 1. End mapping revealed that the novel transcript was transcribed in cis from the strand complementary to rseC and overlapped the entire rseC mRNA. We therefore named it AsrC. From the entire sequence of the novel asRNA, no obvious ORF structure longer than 150 nt or SD element was found. Thus, we hypothesized that the novel asRNA was an ncRNA.

Bottom Line: We found that AsrC increased the levels of rseC mRNA and protein.The expression of rpoE was also increased in S. typhi after overexpression of AsrC, which was dependent on rseC.Thus, we propose that AsrC increased RseC level and indirectly activating RpoE which can initiate fliA expression and promote the motility of S. typhi.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Jiangsu University - School of Medicine Zhenjiang, China ; Danyang People's Hospital of Jiangsu Province Danyang, China.

ABSTRACT
Bacterial non-coding RNAs are essential in many cellular processes, including response to environmental stress, and virulence. Deep sequencing analysis of the Salmonella enterica serovar typhi (S. typhi) transcriptome revealed a novel antisense RNA transcribed in cis on the strand complementary to rseC, an activator gene of sigma factor RpoE. In this study, expression of this antisense RNA was confirmed in S. typhi by Northern hybridization. Rapid amplification of cDNA ends and sequence analysis identified an 893 bp sequence from the antisense RNA coding region that covered all of the rseC coding region in the reverse direction of transcription. This sequence of RNA was named as AsrC. After overexpression of AsrC with recombinantant plasmid in S. typhi, the bacterial motility was increased obviously. To explore the mechanism of AsrC function, regulation of rseC and rpoE expression by AsrC was investigated. We found that AsrC increased the levels of rseC mRNA and protein. The expression of rpoE was also increased in S. typhi after overexpression of AsrC, which was dependent on rseC. Thus, we propose that AsrC increased RseC level and indirectly activating RpoE which can initiate fliA expression and promote the motility of S. typhi.

No MeSH data available.