<i>Escherichia coli</i> inner membrane protein YciB interacts with ZipA that is important for cell division

Badaluddin, Noor Afiza; Kitakawa, Madoka

doi:10.1111/gtc.12299

Cited by 4 publications

(6 citation statements)

References 38 publications

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“…However, we observed the sensitivity to osmotic shock of Δ yciB mutant only in the logarithmic growth phase and not in stationary culture, which suggests that YciB does not participate in the kinetic activity of PG synthesis as expected from its structure, but probably acts by coordinating PG synthesis with cell elongation and cell division to maintain the integrity of the cell envelope. Recently, we found that the septum‐site localization of ZipA, a protein essential for cell division , is disturbed in the Δ yciB mutant , which supports the above postulate about the function of YciB.…”

supporting

confidence: 76%

“…), we C‐terminally fused the T18 fragment of CyaA to YciB (YciB‐T18) on the plasmid pUT18 and appended T25 to the cytoplasmic region of each cell elongation protein on the plasmid pKT25 or pKnT25 (Table S1). Because overproduction of full length YciB is harmful to cells and severely inhibits their growth , we also constructed YciB 170 ‐T18, which lacks C‐terminal 9 amino acid residues of YciB, and examined the interaction. Analysis of fusion proteins revealed that the amount of YciB‐T18 in the cell was about one third of YciB 170 ‐T18 (Fig.…”

mentioning

confidence: 99%

“…Therefore, to fully investigate the nature of the interaction, it is necessary to further examine it by another method. Recently, we found that YciB is localized through the cell along the envelope and interacts directly with ZipA . ZipA is a component of the divisome and observed mostly at the cell division site.…”

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confidence: 99%

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Characterization of inner membrane protein YciB in Escherichia coli: YciB interacts with cell elongation and division proteins

Badaluddin

Kitakawa

2015

Microbiology and Immunology

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The function of inner membrane protein YciB in Escherichia coli has not been identified. In this study, the membrane topology of the protein that contains five transmembrane domains was clarified. YciB was found to interact with various proteins involved in cell elongation and cell division using a bacterial two-hybrid system. It was also found that the deletion mutant of yciB is susceptible to the low osmolarity. These observations together with previous reports indicate that YciB is involved in synthesis of the cell envelope by interacting with cell elongation and cell division complexes.Key words Escherichia coli, membrane topology, two-hybrid analysis, YciB.Bacterial cell envelopes that consist of an IM and the cell wall are the principal stress-bearing and shape-maintaining elements of cells; their integrity is of critical importance to cell viability. The E. coli genome contains over 800 genes (1) that encode IM proteins; however, many of them have not yet been well characterized and their functions remain unknown. We recently identified that one such gene, yciB, is a gene that is required for normal biofilm formation (2) and interacts genetically with rodZ, a gene that is important for maintaining rodtype morphology (3-5). These findings suggest that YciB has a role in cell envelope synthesis.YciB is predicted to be a multi-pass IM protein with five transmembrane domains (Fig. 1a); however, this prediction has not experimentally been examined. Therefore, we first investigated the membrane topology of YciB using a dual pho-lac reporter system (6). In this system, the color on the indicator plate of E. coli strain DH5a (phoA -lacZ -DM15) carrying the pKTop plasmid shows whether the C-terminus of the protein expressed from the cloned gene resides in the cytoplasm or in the periplasm. According to the predicted structure of YciB, we designed serial mutants with deletions from the Cterminus, as shown in Figure 1a, and cloned each fragment into pKTop. The resultant pKTop clones (Table S1) were introduced into DH5a and the topology of the fusion proteins analyzed. Transformants expressing YciB 147 and YciB 75 fusion proteins exhibited blue color, indicating strong phosphatase but low b-galactosidase activity, which indicates that the C-termini of these positions are localized on the periplasmic side of the inner membrane. In contrast, full length YciB, YciB 118 , YciB 48 fusion proteins showed high b-galactosidase activity but low phosphatase activity (red color), showing that the C-termini of these proteins are located in the cytoplasm (Fig. 1b). We also constructed a pKTop recombinant expressing YciB 42-179 fusion. This fusion protein lacks the N-terminal region that is predicted to be periplasmic and the first transmembrane domain. We found that the C-terminus of this mutant is cytoplasmic, as is the full length YciB. Similarly, YciB 42-147 fusion that we deleted at both the N-and C-termini gave the same topological result (blue color) as YciB 147 . These results suggest that neither the N-terminal no...

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confidence: 76%

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confidence: 99%

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Characterization of inner membrane protein YciB in Escherichia coli: YciB interacts with cell elongation and division proteins

Badaluddin

Kitakawa

2015

Microbiology and Immunology

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“…). An earlier study showed reduced growth and shorter cell lengths in cells deleted for yciB , and modest cell elongation upon YciB overexpression (Badaluddin and Kitakawa, ). We note that a different strain background and plasmid were used in that report and may contribute to this discrepancy.…”

Section: Resultsmentioning

confidence: 96%

“…Diverse yciB ‐related phenotypes have been described in the literature: cell separation defects, interaction with division and morphogenetic proteins, biofilm deficiency, resistance to the antibiotic bicyclomycin, requirement in the entry of contact‐dependent inhibition toxins, and colicin cytotoxicity (Mac Siomoin et al ., ; Niba et al ., ; Sharma et al ., ; Lycklama and Driessen, ; Niba et al ., ; Li et al ., ; Willett et al ., ; Badaluddin and Kitakawa, ). Our results of YciB playing a critical role in the barrier function of the OM and OM protein assembly perhaps by modulating a fundamental property of the IM such as PMF maintenance, offer a simplifying explanation for the pleiotropic phenotypes exhibited by yciB mutants described in earlier studies.…”

Section: Discussionmentioning

confidence: 99%

A synergistic role for two predicted inner membrane proteins of Escherichia coli in cell envelope integrity

Mychack

Amrutha

Chung

et al. 2018

Molecular Microbiology

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The bacterial cytoplasmic membrane is a principal site of protein translocation, lipid and peptidoglycan biogenesis, signal transduction, transporters and energy generating components of the respiratory chain. Although 25-30% of bacterial proteomes consist of membrane proteins, a comprehensive understanding of their influence on fundamental cellular processes is incomplete. Here, we show that YciB and DcrB, two small cytoplasmic membrane proteins of previously unknown functions, play an essential synergistic role in maintaining cell envelope integrity of Escherichia coli. Lack of both YciB and DcrB results in pleiotropic cell defects including increased levels of lipopolysaccharide, membrane vesiculation, dynamic shrinking and extension of the cytoplasmic membrane accompanied by lysis and cell death. The stalling of an abundant outer membrane lipoprotein, Lpp, at the periplasmic face of the inner membrane leads to lethal inner membrane-peptidoglycan linkages. Additionally, the periplasmic chaperone Skp contributes to yciB dcrB mutant cell death by possibly mistargeting stalled porins into the inner membrane. Consistent with the idea of a compromised envelope in the yciB dcrB mutant, multiple envelope stress response systems are induced, with Cpx signal transduction being required for growth. Taken together, our results suggest a fundamental role for YciB and DcrB in cell envelope biogenesis.

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Bacterial division FtsZ forms liquid condensates with nucleoid-associated Z-ring inhibitor SlmA

Monterroso

Zorrilla²,

Sobrinos-Sanguino

et al. 2018

Preprint

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17Macromolecular condensation resulting from biologically regulated liquid-liquid phase 18 transitions is emerging as a mechanism to organize the intracellular space in 19 eukaryotic systems, with broad implications in cell physiology and pathology. Here we 20 show that FtsZ, central element of the division ring in most bacteria, forms condensates 21 when in complex with SlmA, the protein preventing septal ring assembly nearby the 22 chromosome in E. coli. The formation of condensates is promoted by crowding and 23 enhanced by sequence-specific binding of SlmA to DNA. These structures are dynamic 24 and FtsZ within them remains active for GTP-triggered fiber formation. Their location is 25 sensitive to compartmentalization and to the presence of a membrane boundary in 26 microfluidics-based cell mimetic systems, likely affecting their reactivity. We propose 27 that reversible condensation may play a role in the modulation of FtsZ assembly and/or 28 location by SlmA and, hence, in the regulation of ring stability, constituting a singular 29 example of a prokaryotic nucleoprotein complex exhibiting this kind of phase transition.30 31 32 33 34 35 36 Recent studies evidence the relevance of the transient condensation of proteins and 37other molecules into structures behaving as liquid droplets in the spatiotemporal 38 regulation of biological processes [1]. The emergence of these membraneless 39 compartments can be described as a liquid demixing process generating phase 40 boundaries to temporarily confine specific functional entities [2]. The selective 41 accumulation of biomolecules in these dynamic compartments may deeply influence 42 their reactivity, enhancing and accelerating their mutual recognition while disfavoring 43 interactions with excluded elements [3][4][5]. Crowding due to the overall high 44 concentration of macromolecules in the cells clearly provides a non-specific driving 45 force favoring phase transitions and condensation [6-10], another contributing factor 46 being, it seems, multivalency [1, 3, 11, 12]. Indeed, most biomolecular condensates 47 analyzed consist of various molecules containing multiple homo or heteroassociation 48 elements, like nucleic acids, usually RNA, and proteins harboring various domains of 49 interaction [1]. The presence of unstructured regions in proteins also seems to promote 50 condensation and a number of intrinsically disordered proteins have been found to form 51 liquid droplets on their own under crowding conditions in vitro and in vivo that evolve 52 towards solid aggregates [9, 10,[13][14][15]. Specific features shared by these 53 condensates, like the ability to exchange molecules with the surroundings, their local 54 and reversible generation in response to changes in component interactions and/or 55 concentrations, and their evolving physical properties [1] make them particularly 56 suitable for the fine tuning of molecular localization and reactivity. 57 Phase separation has been shown to occur in a variety of signaling and other systems 58 in eukaryotes (s...

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Escherichia coli inner membrane protein YciB interacts with ZipA that is important for cell division

Cited by 4 publications

References 38 publications

Characterization of inner membrane protein YciB in Escherichia coli: YciB interacts with cell elongation and division proteins

Characterization of inner membrane protein YciB in Escherichia coli: YciB interacts with cell elongation and division proteins

A synergistic role for two predicted inner membrane proteins of Escherichia coli in cell envelope integrity

Bacterial division FtsZ forms liquid condensates with nucleoid-associated Z-ring inhibitor SlmA

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