• 한국미생물·생명공학회지
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• 제40권1호
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• pp.70-75
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• 2012

살모넬라(Salmonella)의 염색체에 존재하는 병원성 유전자의 집합체인 Salmonella pathogenicity island(SPI)1 과 2는 살모넬라가 유발하는 다양한 질병에 중요한 역할을 한다. SPI1의 발현을 유도하는 HilD는 Luria-Bertani(LB) 배지 조건에서 SPI2의 발현 활성인자로 작용하는 것으로 알려져 있으나 LB 배지 내에서 hilD 유전자의 발현 양상은 아직까지 연구되지 않았다. 본 연구에서는 LB 배지에 살모넬라를 배양하면서 hilD 유전자의 발현과 단백질 양을 조사하였으며 SPI2 유전자인 sseA의 발현과 비교하였다. hilD의 발현은 대수 증식기 경과 후 정지기(stationary phase)로 전환되는 시기에 비약적으로 증가하였으나 sseA의 발현은 정지기 후반부에 최대로 증가하였다. 즉, 후반 정지기에서 HilD 단백질은 낮은 수준으로 존재함에도 불구하고 SPI2의 발현을 유도한다는 것을 알 수 있었다. SPI1의 다른 발현 조절인자인 hilA와 invF의 변이체에서 sseA의 발현을 살펴본 결과 invF의 변이는 hilD와는 다르게 배지 조건에 상관없이 오히려 sseA의 발현을 증가시켰다. 또한, InvF의 과발현은 sseA 발현을 정상 수준으로 복원시켰지만 추가적인 감소는 일으키지 않는다는 것을 알 수 있었다. SPI1은 HilD를 이용하여 SPI2의 발현을 유도하지만 반대로 InvF를 이용하여 발현을 억제하기도 하는 이중적인 조절 기전을 가지고 있는 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1664-1672
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• 2013

In Salmonella enterica serovar Typhimurium, many genes encoded within Salmonella pathogenicity island 1 (SPI1) are required to induce intestinal/diarrheal disease. In this study, we compared the expression of four SPI1 genes (hilA, invF, prgH, and sipC) under shaking and standing culture conditions and found that the expression of these genes was highest during the transition from the exponential to stationary phase under shaking conditions. To identify regulators associated with the stationary phase-dependent activation of SPI1, the effects of selected regulatory genes, including relA/spoT (ppGpp), luxS, ihfB, hfq, and arcA, on the expression of hilA and invF were compared under shaking conditions. Mutations in the hfq and arcA genes caused a reduction in hilA and invF expression (more than 2-fold) in the early stationary phase only, whereas the lack of ppGpp and IHF decreased hilA and invF gene expression during the entire stationary phase. We also found that hfq and arcA mutations caused a reduction of hilD expression upon entry into the stationary phase under shaking culture conditions. Taken together, these results suggest that Hfq and ArcA regulate the hilD promoter, causing an accumulation of HilD, which can trigger a stationary phase-dependent activation of SPI1 genes under shaking culture conditions.

• Journal of Microbiology
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• 제43권spc1호
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• pp.85-92
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• 2005

An early step in the pathogenesis of non-typhoidal Salmonella species is the ability to penetrate the intestinal epithelial monolayer. This process of cell invasion requires the production and transport of secreted effector proteins by a type III secretion apparatus encoded in Salmonella pathogenicity island I (SPI-1). The control of invasion involves a number of genetic regulators and environmental stimuli in complex relationships. SPI-1 itself encodes several transcriptional regulators (HilA, HilD, HilC, and InvF) with overlapping sets of target genes. These regulators are, in turn, controlled by both positive and regulators outside SPI-1, including the two-component regulators BarA/SirA and PhoP/Q, and the csr post-transcriptional control system. Additionally, several environmental conditions are known to regulate invasion, including pH, osmolarity, oxygen tension, bile, $Mg^{2+}$ concentration, and short chain fatty acids. This review will discuss the current understanding of invasion control, with emphasis on the interaction of environmental factors with genetic regulators that leads to productive infection.

• 미생물학회지
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• 제34권3호
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• pp.108-114
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• 1998

Salmonella typhimurium에서 병독성 발현, 특히 숙주세포내로의 침입에 중요한 역할을 하는 유전자들의 집단(gene cluster)인 Salmonella Pathogenicity Island 2(SPI2)의 발현에 대한 다양한 환경요인들의 영향을 조사하였다. 이를 위해 SPI2의 주요 유전자인 ssaJ와 ssaK의 promoter를 포함하는 regulatory region을 promoterless lac operon과 융합시켜 reporter를 제조하였다. 그리고 산소농도, 삼투압, pH, 탄소원 결핍 및 glycerol 참가 등 여러 환경요인들의 변화가 이 reporter 유전자의 발현에 미치는 효과를 조사한 결과 저산소, 낮은 삼투압, 약 알칼리 등이 ssaJ와 ssaK의 발현을 증가시켰으며 위의 세조건이 함께 존재할 때 그 발현이 가장 크게 증가하는 것으로 나타났다. 그러나 탄소원 결핍이나 glycerol 첨가는 이 두 유전자들의 발현에 영향을 주지 않았다. 또한 이상과 같은 환경인자들의 효과는 S. typhimurium의 세 가지 야생형인 LT2, UK1과 SL1344 모두에서 동일한 양상을 보였다. 다른 한편 SPI1의 transcriptional activator를 암호화하는 조절유전자인 hilA의 돌연변이는 ssaJ와 ssaK의 발현에 영향을 미치지 않음도 밝혀냈다. 따라서 이상의 결과는 SPI1과 SPI2가 서로 별개의 조절계에 의해 그 발현이 조절됨을 보여준다.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.175-182
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• 2005

Abstract Salmonella pathogenicity island 1 (SPI1) gene expression is regulated by many environmental signals such as oxygen, osmolarity, and pH. Here, we examined changes in the expression level of various regulatory proteins encoded within SPI1 in response to three different concentrations of NaCl, using primer extension analysis. Transcription of all the regulatory genes tested was activated most when Salmonella were grown in Luria Broth (LB) containing 0.17 M NaCl. The expression of hilA, invF, and hilD was decreased in the presence of 0.47 M NaCl or in the absence of NaCl, while hilC expression was almost constant regardless of the NaCl concentration when Salmonella were grown to exponential phase under low-oxygen condition. The reduced expression of hilA, invF, and hilD resulted in lower invasion of hilC mutant to the cultured animal cells when the mutant was grown in the presence of 0.47 M NaCl or in the absence of NaCl prior to infection. Among the proteins secreted via the SPI1-type III secretion system (TTSS), the level of sopE2 expression was not influenced by medium osmolarity. Various effects of osmolarity on virulence gene regulation observed in this study is one example of multiple regulatory pathways used by Salmonella to cause infection.

• Molecules and Cells
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• 제28권4호
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• pp.389-395
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• 2009

We previously observed that the transcription of some flagellar genes decreased in Salmonella Typhimurium tdcA mutant, which is a gene encoding the transcriptional activator of the tdc operon. Since flagella-mediated bacterial motility accelerates the invasion of Salmonella, we have examined the effect of tdcA mutation on the invasive ability as well as the flagellar biosynthesis in S. Typhimurium. A tdcA mutation caused defects in motility and formation of flagellin protein, FliC in S. Typhimurium. Invasion assays in the presence of a centrifugal force confirmed that the defect of flagellum synthesis decreases the ability of Salmonella to invade into cultured epithelial cells. In addition, we also found that the expression of Salmonella pathogenicity island 1 (SPI1) genes required for Salmonella invasion was down-regulated in the tdcA mutant because of the decreased expression of fliZ, a positive regulator of SPI1 transcriptional activator, hilA. Finally, the virulence of a S. Typhimurium tdcA mutant was attenuated compared to a wild type when administered orally. This study implies the role of tdcA in the invasion process of S. Typhimurium.

• Journal of Microbiology and Biotechnology
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• 제28권11호
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• pp.1896-1907
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• 2018

Salmonellosis is commonly associated with meat and poultry products, but an increasing number of Salmonella outbreaks have been attributed to contaminated vegetables and fruits. Enteric pathogens including Salmonella enterica spp. can colonize diverse produce and persist for a long time. Considering that fresh vegetables and fruits are usually consumed raw without heat treatments, Salmonella contamination may subsequently lead to serious human infections. In order to understand the underlying mechanism of Salmonella adaptation to produce, we investigated the transcriptomics of Salmonella in contact with green vegetables, namely cabbage and napa cabbage. Interestingly, Salmonella pathogenicity island (SPI)-1 genes, which are required for Salmonella invasion into host cells, were up-regulated upon contact with vegetables, suggesting that SPI-1 may be implicated in Salmonella colonization of plant tissues as well as animal tissues. Furthermore, Salmonella transcriptomic profiling revealed several genetic loci that showed significant changes in their expression in response to vegetables and were associated with bacterial adaptation to unfavorable niches, including STM14_0818 and STM14_0817 (speF/potE), STM14_0880 (nadA), STM14_1894 to STM14_1892 (fdnGHI), STM14_2006 (ogt), STM14_2269, and STM14_2513 to STM14_2523 (cbi operon). Here, we show that nadA was required for bacterial growth under nutrient-restricted conditions, while the other genes were required for bacterial invasion into host cells. The transcriptomes of Salmonella in contact with cabbage and napa cabbage provided insights into the comprehensive bacterial transcriptional response to produce and also suggested diverse virulence determinants relevant to Salmonella survival and adaptation.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1729-1738
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• 2020

Salmonellosis is a form of gastroenteritis caused by Salmonella infection. The main transmission route of salmonellosis has been identified as poorly cooked meat and poultry products contaminated with Salmonella. However, in recent years, the number of outbreaks attributed to contaminated raw produce has increased dramatically. To understand how Salmonella adapts to produce, transcriptomic analysis was conducted on Salmonella enterica serovar Virchow exposed to fresh-cut radish greens. Considering the different Salmonella lifestyles in contact with fresh produce, such as motile and sessile lifestyles, total RNA was extracted from planktonic and epiphytic cells separately. Transcriptomic analysis of S. Virchow cells revealed different transcription profiles between lifestyles. During bacterial adaptation to fresh-cut radish greens, planktonic cells were likely to shift toward anaerobic metabolism, exploiting nitrate as an electron acceptor of anaerobic respiration, and utilizing cobalamin as a cofactor for coupled metabolic pathways. Meanwhile, Salmonella cells adhering to plant surfaces showed coordinated upregulation in genes associated with translation and ribosomal biogenesis, indicating dramatic cellular reprogramming in response to environmental changes. In accordance with the extensive translational response, epiphytic cells showed an increase in the transcription of genes that are important for bacterial motility, nucleotide transporter/metabolism, cell envelope biogenesis, and defense mechanisms. Intriguingly, Salmonella pathogenicity island (SPI)-1 and SPI-2 displayed up- and downregulation, respectively, regardless of lifestyles in contact with the radish greens, suggesting altered Salmonella virulence during adaptation to plant environments. This study provides molecular insights into Salmonella adaptation to plants as an alternative environmental reservoir.