Abstract
Antisense RNAs from complementary strands of protein coding genes regulate the expression of genes involved in many cellular processes. Using deep sequencing analysis of the Salmonella enterica serovar Typhi (S. Typhi) transcriptome, a novel antisense RNA encoded on the strand complementary to the rpoH gene was revealed. In this study, the molecular features of this antisense RNA were assessed using northern blotting and rapid amplification of cDNA ends. The 3,508 nt sequence of RNA was identified as the antisense RNA of the rpoH gene and was named ArpH. ArpH was found to attenuate the invasion of HeLa cells by S. Typhi by regulating the expression of SPI-1 genes. In an rpoH mutant strain, the invasive capacity of S. Typhi was increased, whereas overexpression of ArpH positively regulates rpoH mRNA levels. Results of this study suggest that the cis-encoded antisense RNA ArpH is likely to affect the invasive capacity of S. Typhi by regulating the expression of rpoH.
Highlights
We described the identification and characterization of a novel antisense RNA encoded by the strand complementary to the rpoH gene sequence that we named ArpH
We found that the arpH fragment was approximately 3,000 nt (Figure 1B). 5 -RACE was performed to map the 5 end of the arpH sequence, which was found to be 411 nt upstream of the yhhK initiation codon (Figure 1A). 3 -RACE revealed that the 3 -end of the arpH sequence was 238 nt upstream of the rpoH initiation codon on the complementary strand
To examine the possibility that ArpH may play a role in the invasion of epithelial cells by Salmonella, we examined the invasion efficiency of wild-type, arpH, and rpoH strains into HeLa cells
Summary
Typhi to invade and survive in non-phagocytic cells is an indication of its pathogenicity These pathogenic mechanisms have been linked to SPIs (Nieto et al, 2016). Pathogens can quickly adapt to changing circumstances by regulating expression of their virulence genes during the process of infection. Many non-coding RNAs (ncRNAs) involved in complex regulatory mechanisms have been identified. Trans-encoded ncRNAs are located between protein coding genes and, because they are located at distant genomic locations, are only partially complementary to their target mRNAs and act via incomplete base pairing. RpoH activates expression of the hfq gene, which encodes an RNA-binding regulatory protein. We described the identification and characterization of a novel antisense RNA encoded by the strand complementary to the rpoH gene sequence that we named ArpH.
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