Biology subjects

Slauch, J. M.

Publications and source records attributed to Slauch, J. M..

5 recordsLinked to original sources

bioRxiv2026

CorB and MgpB (YegH) alleviate excess-magnesium stress and contribute to egg white tolerance and virulence in Salmonella

Maintaining proper cation homeostasis is essential for the survival of all living organisms. During infection, Salmonella must adapt to host-imposed magnesium (Mg2+) limitation. To compensate, Salmonella induces high-affinity Mg2+ transporters and synthesis of polyamines, organic cations that partially substitute for Mg2+. Once Mg2+ levels are restored and cells enter stationary phase, excess cations must be exported; failure to do so results in loss of viability. The CorC-domain protein PaeA exports excess polyamines, whereas MgpA and CorC reduce intracellular Mg2+ levels. Here, we identify two new CorC-domain proteins, MgpB (formerly YegH) and CorB, as mediators of Mg2+ homeostasis, showing that they also reduce intracellular Mg2+ accumulation and protect cells from endogenous excess-cation stress. Genetic analyses revealed the functional redundancy among these proteins in conferring resistance to exogenous high Mg2+, endogenous stress induced after Mg2+ starvation, and egg whites. Deletion of mgpB and/or corB conferred little to no phenotype. However, these mutations showed synergistic phenotypes in strains lacking MgpA. This synergistic relationship was also evident in mouse competition assays. Lack of MgpA, CorC, MgpB, and CorB leads to an increase in intracellular Mg2+ levels. Moreover, expression of mgpB and corB are increased in high Mg2+, all consistent with their role as putative Mg2+ exporters. Together, these findings identify MgpB and CorB as new mediators of Mg2+ homeostasis and reveal a multilayered system that enables Salmonella to finely balance intracellular cation levels during environmental and host-imposed stress.

microbiology↗

bioRxiv2025

The CorC proteins MgpA (YoaE) and CorC protect from excess-cation stress and are required for egg white tolerance and virulence in Salmonella

Cation homeostasis is a vital function. In Salmonella, growth in very low Mg2+ induces expression of high-affinity Mg{superscript 2} transporters and synthesis of polyamines, organic cations that substitute for Mg{superscript 2}. Once Mg{superscript 2} levels are re-established, the polyamines must be excreted by PaeA. Otherwise, cells lose viability due to a condition we term excess-cation stress. We sought additional tolerance mechanisms for this stress. We show that CorC and MgpA (YoaE) are essential for survival in stationary phase after Mg2+ starvation. Deletion of corC causes a loss of viability additive with the paeA phenotype. Deletion of mgpA causes a synthetic defect in the corC background. This lethality is suppressed by loss of the inducible Mg2+ transporters, suggesting that the corC mgpA mutant is sensitive to changes in intracellular Mg2+. CorC and MgpA function independently of PaeA. A paeA mutant is sensitive to externally added polyamine in stationary phase; loss of CorC and MgpA suppressed this sensitivity. Conversely, the corC mgpA mutant, but not the paeA mutant, exhibited sensitivity to high Mg2+ and egg white. The corC mgpA mutant is also attenuated in a mouse model. The corC and mgpA genes are induced in response to increased Mg2+ concentrations. Thus, CorC and MgpA play some interrelated role in cation homeostasis. It is unlikely that these phenotypes are due to absolute levels of cations. Rather, the cell maintains relative concentrations of various cations that likely compete for binding to anionic components. Imbalance of these cations affects some essential function(s), leading to a loss of viability. IMPORTANCEMg{superscript 2} and other cations are critical for counteracting anionic compounds in the cell including RNA, DNA, and nucleotides. Both excessively low and high cation levels are toxic. To maintain proper intracellular concentrations, cells must regulate Mg{superscript 2} importers and exporters, or modulate the levels of other cations or anions that affect free Mg{superscript 2} levels. In Salmonella, no mutants sensitive to high Mg{superscript 2} levels have been identified. Here, we demonstrate that the largely uncharacterized proteins MgpA and CorC are induced under high Mg{superscript 2} conditions and are essential for tolerance to high Mg{superscript 2} levels. These genes are also essential for survival during endogenous excess-cation stress triggered by the transition to stationary phase after Mg{superscript 2} starvation, as well as for virulence, highlighting the broader role of cation homeostasis.

microbiology↗

bioRxiv2024

The Salmonella pathogenicity island 1-encoded small RNA InvR mediates post-transcriptional feedback control of the activator HilA in Salmonella

Salmonella Pathogenicity Island 1 (SPI1) encodes a type three secretion system (T3SS) essential for Salmonella invasion of intestinal epithelial cells. Many environmental and regulatory signals control SPI1 gene expression, but in most cases, the molecular mechanisms remain unclear. Many of these regulatory signals control SPI1 at a post-transcriptional level and we have identified a number of small RNAs (sRNAs) that control the SPI1 regulatory circuit. The transcriptional regulator HilA activates expression of the genes encoding the SPI1 T3SS structural and primary effector proteins. Transcription of hilA is controlled by the AraC-like proteins HilD, HilC, and RtsA. The hilA mRNA 5 untranslated region (UTR) is ~350-nuclotides in length and binds the RNA chaperone Hfq, suggesting it is a likely target for sRNA-mediated regulation. We used the rGRIL-seq (reverse global sRNA target identification by ligation and sequencing) method to identify sRNAs that bind to the hilA 5 UTR. The rGRIL-seq data, along with genetic analyses, demonstrate that the SPI1-encoded sRNA InvR base pairs at a site overlapping the hilA ribosome binding site. HilD and HilC activate both invR and hilA. InvR in turn negatively regulates the translation of the hilA mRNA. Thus, the SPI1-encoded sRNA InvR acts as a negative feedback regulator of SPI1 expression. Our results suggest that InvR acts to fine-tune SPI1 expression and prevent overactivation of hilA expression, highlighting the complexity of sRNA regulatory inputs controlling SPI1 and Salmonella virulence. IMPORTANCESalmonella Typhimurium infections pose a significant public health concern, leading to illnesses that range from mild gastroenteritis to severe systemic infection. Infection is initiated and requires a complex apparatus that the bacterium uses to invade the intestinal epithelium. Understanding how Salmonella regulates this system is essential for addressing these infections effectively. Here we show that the small RNA (sRNA) InvR imposes negative feedback regulation on expression of the invasion system. This work underscores the role of sRNAs in Salmonellas complex regulatory network, offering new insights into how these molecules contribute to bacterial adaptation and pathogenesis.

microbiology↗

bioRxiv2022

Small RNAs activate Salmonella pathogenicity island 1 by modulating mRNA stability through the hilD mRNA 3' UTR

Salmonella enterica serovar Typhimurium is an enteric pathogen associated with food-borne disease. Salmonella invades the intestinal epithelium using a type three secretion system encoded in Salmonella pathogenicity Island 1 (SPI-1). SPI-1 genes are tightly regulated by a complex feed-forward loop to ensure proper spatial and temporal expression. Most regulatory input is integrated at HilD, through control of hilD mRNA translation or HilD protein activity. The hilD mRNA possesses a 310-nucleotide 3' untranslated region (UTR) that influences HilD and SPI-1 expression, and this regulation is dependent on Hfq and RNase E, cofactors known to mediate small RNA (sRNA) activities. Thus, we hypothesized that the hilD mRNA 3' UTR is a target for sRNAs. Here we show that the sRNAs, SdsR and Spot 42 regulate SPI-1 by targeting different regions of the hilD mRNA 3' UTR. Regulatory activities of these sRNAs depend on Hfq and RNase E, in agreement with previous roles found for both at the hilD 3' UTR. We show that SdsR and RNase E are responsible for the accumulation of variable fragments of the hilD mRNA 3' UTR. Collectively, this work suggests that these sRNAs targeting the hilD mRNA 3' UTR regulate hilD mRNA levels by interfering with RNase E-dependent mRNA degradation. Our work provides novel insights into mechanisms of sRNA regulation at bacterial mRNA 3' UTRs and adds to our knowledge of post-transcriptional regulation of the SPI-1 complex feed-forward loop. ImportanceSalmonella are prominent food-borne pathogens, infecting millions of people a year. To express virulence genes at the correct time and place in the host, Salmonella uses a complex regulatory network that senses environmental conditions. Known for their role in allowing quick responses to stress and virulence conditions, we investigate the role of small RNAs in facilitating precise expression of these genes. We provide evidence that the 3' untranslated region of the hilD mRNA, encoding a key virulence regulator, is a target for small RNAs and the ribonuclease RNase E. The small RNAs play a role in stabilizing hilD mRNA to allow proper expression of Salmonella virulence genes in the host.

microbiology↗

bioRxiv2021

The sRNA MicC downregulates hilD translation to control the SPI1 T3SS in Salmonella enterica serovar Typhimurium

Salmonella enterica serovar Typhimurium invades the intestinal epithelium and induces inflammatory diarrhea using the Salmonella pathogenicity island 1 (SPI1) type III secretion system (T3SS). Expression of the SPI1 T3SS is controlled by three AraC-like regulators, HilD, HilC and RtsA, which form a feed-forward regulatory loop that leads to activation of hilA, encoding the main transcriptional regulator of the T3SS structural genes. This complex system is affected by numerous regulatory proteins and environmental signals, many of which act at the level of hilD mRNA translation or HilD protein function. Here, we show that the sRNA MicC blocks translation of the hilD mRNA by base pairing near the ribosome binding site. This binding blocks translation but does not induce degradation of the hilD message. Our data indicate that micC is transcriptionally activated by SlyA, and SlyA feeds into the SPI1 regulatory network solely through MicC. Transcription of micC is negatively regulated by the OmpR/EnvZ two-component system, but this regulation is dependent on SlyA. OmpR/EnvZ control SPI1 expression partially through MicC, but also affect expression through other mechanisms. MicC-mediated regulation plays a role during infection, as evidenced by an increase in Salmonella fitness in the intestine in the micC deletion mutant that is dependent on the SPI1 T3SS. These results further elucidate the complex regulatory network controlling SPI1 expression and add to the list of sRNAs that control this primary virulence factor. IMPORTANCEThe Salmonella SPI1 T3SS is the primary virulence factor required for causing intestinal disease and initiating systemic infection. The system is regulated in response to a large variety of environmental and physiological factors such that the T3SS is expressed at only the appropriate time and place in the host during infection. Here we show how the sRNA MicC affects expression of the system. This work adds to our detailed mechanistic studies aimed at a complete understanding of the regulatory circuit.

microbiology↗