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Yoon, Ki-Hong The Korean Society for Applied Biological Chemistr 2005 Journal of Applied Biological Chemistry (J. Appl. Vol.48 No.4
Nucleotide sequence of Brevibacterium flavum ptsG gene capable of complementing Escherichia coli ZSC113 mutations defective to glucose permease activity of phosphotransferase system was completely determined, and the gene product was compared with other glucose-specific enzyme II ($EII^{Glc}$). A ptsG gene of B. flavum consisted of open reading frame of 2,025 nucleotides putatively encoding polypeptide of 675 amino acid residues and TAA stop codon. Deduced amino acid sequence of B. flavum ($EII^{Glc}$) had high homology with ($EIIs^{Glc}$) of Corynebacterium glutamicum, C. efficiens, and B. lactofermentum. Arrangement of structural domains, IIBCA, of B. flanum ($EII^{Glc}$) protein was identical to that of EIIs belonging to glucose-phosphotransferase system.
Lee, Jae-Woo,Park, Young-Ha,Seok, Yeong-Jae National Academy of Sciences 2018 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.115 No.29
<▼1><P><B>Significance</B></P><P>Most bacteria accumulate the molecular alarmone (p)ppGpp to divert resources away from growth and division toward biosynthesis under various nutrient limitations. Despite its crucial role, uncontrolled accumulation of this alarmone causes severe growth inhibition and cell death. Thus, fine-tuning the cellular (p)ppGpp level is required to ensure survival and adaptation under harsh nutritional conditions. Here, we identify Rsd as a stimulator of the (p)ppGpp-degrading activity of SpoT during carbon source downshift in <I>Escherichia coli</I>, and this regulation is controlled by the phosphorylation state of HPr, a general component of the PEP-dependent sugar transport system. This study establishes a direct link between sugar signaling and the bacterial stringent response.</P></▼1><▼2><P>Bacteria respond to nutritional stresses by changing the cellular concentration of the alarmone (p)ppGpp. This control mechanism, called the stringent response, depends on two enzymes, the (p)ppGpp synthetase RelA and the bifunctional (p)ppGpp synthetase/hydrolase SpoT in <I>Escherichia coli</I> and related bacteria. Because SpoT is the only enzyme responsible for (p)ppGpp hydrolysis in these bacteria, SpoT activity needs to be tightly regulated to prevent the uncontrolled accumulation of (p)ppGpp, which is lethal. To date, however, no such regulation of SpoT (p)ppGpp hydrolase activity has been documented in <I>E. coli</I>. In this study, we show that Rsd directly interacts with SpoT and stimulates its (p)ppGpp hydrolase activity. Dephosphorylated HPr, but not phosphorylated HPr, of the phosphoenolpyruvate-dependent sugar phosphotransferase system could antagonize the stimulatory effect of Rsd on SpoT (p)ppGpp hydrolase activity. Thus, we suggest that Rsd is a carbon source-dependent regulator of the stringent response in <I>E. coli</I>.</P></▼2>
Yong Zhi,Shunmei Lin,A-Yeung Jang,Ki Bum Ahn,Hyun Jung Ji,Hui-Chen Guo,임상용,Ho Seong Seo 한국미생물학회 2019 The journal of microbiology Vol.57 No.1
Salmonella enterica is a major human pathogen that causes invasive non-typhoidal Salmonellosis (iNTS), resulting in significant morbidity and mortality. Although a number of pre-clinical and clinical studies have reported on the feasibility of developing a safe and effective vaccine against iNTS, there have been no licensed Salmonella vaccines available to protect against NTS strains. Vaccine formulations of highest priority for NTS are live attenuated vaccines, which can elicit effective induction of intestinal mucosal and intracellular bacteria-specific cell mediated immune responses. Since glucose is crucial for intracellular survival and replication in host cells, we constructed strains with mutations in components of the glucose uptake system, called the phosphotransferase system (PTS), and compared the relative virulence and immune responses in mice. In this study, we found that the strain with mutations in both ptsI and crr (KST0556) was the most attenuated strain among the tested strains, and proved to be highly effective in inducing a mucosal immune response that can protect against NTS infections in mice. Thus, we suggest here that KST0556 (ΔptsIΔcrr) is a potential live vaccine candidate for NTS, and may also be a candidate for a live delivery vector for heterologous antigens. Moreover, since PTS is a well-conserved glucose transporter system in both Gramnegative and Gram-positive bacteria, the ptsI and crr genes may be potential targets for creating live bacterial vectors or vaccine strains.
Kwon, Il,Lee, Kyu Nam,Lee, Jung Kee,Pan, Jae Gu,Oh, Tae Kwang,Lee, Hyung Hoan,Yoon, Ki Hong 한국미생물 · 생명공학회 1995 Journal of microbiology and biotechnology Vol.5 No.4
A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSC113 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or mannose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-α-D-glucopyranoside (MeGlc) affected the growth of ZSC113 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSC113 cells without pBFT93. It was also found that both 2-deoxyD-[U^-14C]glucose and methyl-αa-D-[U^-14C]glucopyranoside could be effectively transported into ZSC113 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.
Lee, Jung Kee,Sung, Moon Hee,Yoon, Ki Hong,Pan, Jae Gu,Yu, Ju Hyun,Oh, Tae Kwang 한국미생물 · 생명공학회 1993 Journal of microbiology and biotechnology Vol.3 No.1
The gene for mannose enzyme II of phosphoenolpyruvate-dependent phosphotransferase system from Corynebacterium glutamicum KCTC 1445 was cloned into Escherichia coli ZSC113 using plasmid pBR322. The recombinant plasmid, designated pCTS3, contained 2.2 kb DNA fragment, and the physical map of the cloned DNA fragment was determined. The E. coli ptsM ptsG mutant transformed with pCTS3 restored glucose and mannose fermentation ability, and grew well on these sugars as the sole carbon source in the minimal medium. The transformant harboring pCTS3 showed a PTS-mediated repression of growth on maltose by mannose analogue, 2-deoxyglucose. The specificity of the response to 2DG therefore indicates that the cloned DNA fragment carries mannose enzyme II gene.
이고은,서정용 한국자기공명학회 2015 Journal of the Korean Magnetic Resonance Society Vol.19 No.1
The mannitol transporter Enzyme IIMtl of the bacterial phosphotransferase system has two cytoplasmic phosphoryl transfer domains IIAMtl and IIBMtl. The two domains are linked by a flexible peptide linker in mesophilic bacterial strains, whereas they are expressed as separated domains in thermophilic strains. Here, we carried out backbone assignment of IIAMtl from thermophilic Thermoanaerobacter Tencongensis using a suite of heteronuclear triple resonance NMR spectroscopy. We have completed 94% of the backbone assignment, and obtained secondary structural information based on torsion angles derived from the chemical shifts. IIAMtl of Thermoanaerobacter Tencongensis is predicted to have six b strands and six a helices, which is analogous to IIAMtl of Escherichia coli.
Lim, Sangyoung,Seo, Ho Seong,Jeong, Jisu,Yoon, Hyunjin Elsevier 2019 MICROBIOLOGICAL RESEARCH Vol.223 No.-
<P><B>Abstract</B></P> <P>The phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) catalyzes the translocation of sugar substrates with their concomitant phosphorylation in bacteria. In addition to its intrinsic role in sugar transport and metabolism, numerous recent studies report the versatility of the PTS to interconnect energy and signal transduction in response to sugar availability. In this study, the role of PTS in <I>Salmonella</I> virulence regulation was explored. To decipher the regulatory network coordinated by the PTS during <I>Salmonella</I> infection, a transcriptomic approach was applied to a transposon insertion mutant with defective expression of <I>ptsI</I> and <I>crr</I>, which encode enzyme I and enzyme IIA<SUP>Glc</SUP> of the PTS, respectively. There were 114 differentially expressed genes (DEGs) exhibiting two-fold or higher expression changes in the transposon mutant strain, with 13 up-regulated genes versus 101 down-regulated genes. One-third of the DEGs were associated with energy production and carbohydrate/amino acid metabolism pathways, implicating the prominent role of the PTS in carbohydrate transport. With regard to regulation of virulence, the tested mutant decreased the expression of genes associated with quorum sensing, <I>Salmonella</I> pathogenicity islands, flagella, and the PhoPQ regulon. We investigated the possibility of PTS-mediated regulation of virulence determinants identified in the transcriptomic analysis and proposed a regulatory circuit orchestrated by the PTS in <I>Salmonella</I> infection of host cells. These results suggest that <I>Salmonella</I> divergently controls virulence attributes in accordance with the availability of carbohydrates in the environment.</P>
질소-인산전달계 최종 인산 수용체 EIIANtr 조절의 다면성
박찬서,이은진 한국미생물학회 2022 미생물학회지 Vol.58 No.3
질소-인산전달계(PTSNtr)은 다양한 그람 음성 세균에 존재하는 이차적 인산화 매개 세포 신호 조절 시스템이다. 당-인산전달계(Sugar-PTS)가 탄소 영양분과 그 대사물을 기반으로 세포 내에서 다양한 생리 기능을 하는 것처럼 PTSNtr 또한 균주의생장 환경에 따라 다양한 생리 조절 메커니즘을 갖는다. 최근에는 질소/탄소 대사, K⁺ 농도 조절, 병원성 조절을 포함해서 다양한 PTSNtr의 최종 인산 수용체인 EIIANtr의 세포 전체적인 조절 메커니즘에 대한 연구가 진행되어왔다. 해당 시스템을 보유한 균주의 다양성에 따라, EIIANtr에 의한 조절 작용을 매개로한 다양한 표현형이 보고되었으며, 몇몇 조절 기작의 분자적메커니즘은 아직 밝혀지지 않았다. 본 총설은 특정 환경 조건에 반응하여 다양한 종에서 나타나는 EIIANtr의 조절에 의한 표현형과, 해당 조절 메커니즘에 대한 최근의 발견을 정리한다