Structural characterization of the essential cell division protein FtsE and its interaction with FtsX in Streptococcus pneumoniae

ABSTRACTFtsEX is a membrane complex widely conserved across diverse bacterial genera and involved in critical processes such as recruitment of division proteins and in spatial and temporal regulation of muralytic activity during cell division or sporulation. FtsEX is a member of the ABC transporter superfamily, where FtsX is an integral membrane protein and FtsE is an ATPase, required for mechanotransmission of the signal from the cytosol through the membrane, to regulate the activity of cell-wall hydrolases in the periplasm. Both proteins are essential in the major human respiratory pathogenic bacterium, Streptococcus pneumoniae and interact with the modular peptidoglycan hydrolase PcsB at the septum. Here, we report the high-resolution structures of pneumococcal FtsE in complex with different nucleotides. Structural analysis reveals that FtsE contains all the conserved structural motifs associated with ATPase activity, and allowed interpretation of the in vivo dimeric arrangement in both ADP and ATP states. Interestingly, three specific FtsE regions were identified with high structural plasticity that shape the cavity in which the cytosolic region of FtsX would be inserted. The residues corresponding to the FtsX coupling helix, responsible for FtsE contact, were identified and validated by in vivo mutagenesis studies showing that this interaction is essential for cell growth and proper morphology.IMPORTANCEBacterial cell division is a central process that requires exquisite orchestration of both the cell wall biosynthetic and lytic machineries. The essential membrane complex FtsEX, widely conserved across bacteria, play a central role by recruiting proteins to the divisome apparatus and by regulating periplasmic muralytic activity from the cytosol. FtsEX is a member of the Type VII family of the ABC-superfamily but instead transporter, couple ATP hydrolysis by FtsE to mechanically transduce a conformational signal to activate PG hydrolases. So far, no structural information is available for FtsE. Here we provide the structural characterization of FtsE confirming its ATPase nature and revealing regions with high structural plasticity key for FtsX binding. The complementary region in FtsX has been also identified and validated in vivo. Our results provide evidences on how difference between ATP and ADP states in FtsE would dramatically alter FtsEX interaction with PG hydrolase PcsB in pneumococcal division.

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Structural Characterization of the Essential Cell Division Protein FtsE and Its Interaction with FtsX in Streptococcus pneumoniae

mBio ◽

10.1128/mbio.01488-20 ◽

2020 ◽

Vol 11 (5) ◽

Cited By ~ 1

Author(s):

Martin Alcorlo ◽

Daniel Straume ◽

Joe Lutkenhaus ◽

Leiv Sigve Håvarstein ◽

Juan A. Hermoso

Keyword(s):

Cell Wall ◽

Cell Division ◽

Streptococcus Pneumoniae ◽

Structural Characterization ◽

Bound States ◽

Structural Information ◽

Structural Plasticity ◽

Membrane Complex

ABSTRACT FtsEX is a membrane complex widely conserved across diverse bacterial genera and involved in critical processes such as recruitment of division proteins and in spatial and temporal regulation of muralytic activity during cell division or sporulation. FtsEX is a member of the ABC transporter superfamily. The component FtsX is an integral membrane protein, whereas FtsE is an ATPase and is required for the transmission of a conformational signal from the cytosol through the membrane to regulate the activity of cell wall hydrolases in the periplasm. Both proteins are essential in the major human respiratory pathogenic bacterium Streptococcus pneumoniae, and FtsX interacts with the modular peptidoglycan hydrolase PcsB at the septum. Here, we report high-resolution structures of pneumococcal FtsE bound to different nucleotides. Structural analysis revealed that FtsE contains all the conserved structural motifs associated with ATPase activity and afforded interpretation of the in vivo dimeric arrangement in both the ADP and ATP states. Interestingly, three specific FtsE regions with high structural plasticity were identified that shape the cavity in which the cytosolic region of FtsX would be inserted. The residues corresponding to the FtsX coupling helix, responsible for contacting FtsE, were identified and validated by in vivo mutagenesis studies showing that this interaction is essential for cell growth and proper morphology. IMPORTANCE Bacterial cell division is a central process that requires exquisite orchestration of both the cell wall biosynthetic and lytic machineries. The essential membrane complex FtsEX, widely conserved across bacteria, plays a central role by recruiting proteins to the divisome apparatus and by regulating periplasmic muralytic activity from the cytosol. FtsEX is a member of the type VII family of the ABC-superfamily, but instead of being a transporter, it couples the ATP hydrolysis catalyzed by FtsE to mechanically transduce a conformational signal that provokes the activation of peptidoglycan (PG) hydrolases. So far, no structural information is available for FtsE. Here, we provide the structural characterization of FtsE, confirming its ATPase nature and revealing regions with high structural plasticity which are key for FtsE binding to FtsX. The complementary binding region in FtsX has also been identified and validated in vivo. Our results provide evidence on how the difference between the ATP/ADP-bound states in FtsE would dramatically alter the interaction of FtsEX with the PG hydrolase PcsB in pneumococcal division.

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Characterization of divIVA and Other Genes Located in the Chromosomal Region Downstream of the dcw Cluster in Streptococcus pneumoniae

Journal of Bacteriology ◽

10.1128/jb.185.20.6209-6214.2003 ◽

2003 ◽

Vol 185 (20) ◽

pp. 6209-6214 ◽

Cited By ~ 68

Author(s):

Daniela Fadda ◽

Carla Pischedda ◽

Fabrizio Caldara ◽

Michael B. Whalen ◽

Daniela Anderluzzi ◽

...

Keyword(s):

Cell Wall ◽

Cell Division ◽

Streptococcus Pneumoniae ◽

Cell Shape ◽

Chromosomal Region ◽

Chromosome Region ◽

Gene Organization ◽

Transcriptional Analysis ◽

Severe Growth

ABSTRACT We analyzed the chromosome region of Streptococcus pneumoniae located downstream of the division and cell wall (dcw) cluster that contains the homolog of the Bacillus subtilis cell division gene divIVA and some genes of unknown function. Inactivation of divIVA in S. pneumoniae resulted in severe growth inhibition and defects in cell shape, nucleoid segregation, and cell division. Inactivation of the ylm genes resulted in some morphological and/or division abnormalities, depending on the inactivated gene. Transcriptional analysis revealed a relationship between these genes and the ftsA and ftsZ cell division genes, also indicating that the connection between the dcw cluster and the divIVA region is more extensive than just chromosomal position and gene organization.

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Structural characterization of an alkali‐soluble polysaccharide from Angelica sinensis and its antitumor activity in vivo

Chemistry & Biodiversity ◽

10.1002/cbdv.202100089 ◽

2021 ◽

Author(s):

Yan Zhao ◽

Yingying Feng ◽

Xue Jing ◽

Yining Liu ◽

Anjun Liu

Keyword(s):

Antitumor Activity ◽

Structural Characterization ◽

Angelica Sinensis ◽

Soluble Polysaccharide

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In vivo degradation of banana starch: Structural characterization of the degradation process

Carbohydrate Polymers ◽

10.1016/j.carbpol.2010.02.022 ◽

2010 ◽

Vol 81 (2) ◽

pp. 291-299 ◽

Cited By ~ 25

Author(s):

Fernanda H.G. Peroni-Okita ◽

Renata A. Simão ◽

Mateus B. Cardoso ◽

Claudinéia A. Soares ◽

Franco M. Lajolo ◽

...

Keyword(s):

Structural Characterization ◽

Degradation Process ◽

In Vivo Degradation

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Hepatoprotection mechanism against alcohol-induced liver injury in vivo and structural characterization of Pinus koraiensis pine nut polysaccharide

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2020.03.168 ◽

2020 ◽

Vol 154 ◽

pp. 1007-1021

Author(s):

Hang Qu ◽

Xin Gao ◽

Cuilin Cheng ◽

Haitian Zhao ◽

Zhenyu Wang ◽

...

Keyword(s):

Liver Injury ◽

Structural Characterization ◽

Pinus Koraiensis ◽

Pine Nut

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Functional and structural characterization of a novel putative cysteine protease cell wall-modifying multi-domain enzyme selected from a microbial metagenome

Scientific Reports ◽

10.1038/srep38031 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 4

Author(s):

Muhammad Faheem ◽

Diogo Martins-de-Sa ◽

Julia F. D. Vidal ◽

Alice C. M. Álvares ◽

José Brandão-Neto ◽

...

Keyword(s):

Cell Wall ◽

Structural Characterization ◽

Cysteine Protease

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Extracting structural information of Au colloids at ultra-dilute concentrations: identification of growth during nanoparticle immobilization

Nanoscale Advances ◽

10.1039/c9na00159j ◽

2019 ◽

Vol 1 (7) ◽

pp. 2546-2552 ◽

Cited By ~ 1

Author(s):

George F. Tierney ◽

Donato Decarolis ◽

Norli Abdullah ◽

Scott M. Rogers ◽

Shusaku Hayama ◽

...

Keyword(s):

Absorption Spectroscopy ◽

Structural Characterization ◽

Structural Information ◽

Particle Growth ◽

Au Nanoparticle ◽

X Ray ◽

Colloidal Au ◽

X Ray Absorption ◽

Au Colloids

This paper describes the structural characterization of ultra-dilute colloidal Au nanoparticle solutions using X-ray absorption spectroscopy (XAS) and the particle growth during immobilization.

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The in vivo impact of MsLAC1, a Miscanthus laccase isoform, on lignification and lignin composition contrasts with its in vitro substrate preference

BMC Plant Biology ◽

10.1186/s12870-019-2174-3 ◽

2019 ◽

Vol 19 (1) ◽

Author(s):

Feng He ◽

Katja Machemer-Noonan ◽

Philippe Golfier ◽

Faride Unda ◽

Johanna Dechert ◽

...

Keyword(s):

Cell Wall ◽

Transcription Factors ◽

Lignin Content ◽

Lignin Biosynthesis ◽

Lignin Composition ◽

Xylem Fibers ◽

Breeding Target

Abstract Background Understanding lignin biosynthesis and composition is of central importance for sustainable bioenergy and biomaterials production. Species of the genus Miscanthus have emerged as promising bioenergy crop due to their rapid growth and modest nutrient requirements. However, lignin polymerization in Miscanthus is poorly understood. It was previously shown that plant laccases are phenol oxidases that have multiple functions in plant, one of which is the polymerization of monolignols. Herein, we link a newly discovered Miscanthus laccase, MsLAC1, to cell wall lignification. Characterization of recombinant MsLAC1 and Arabidopsis transgenic plants expressing MsLAC1 were carried out to understand the function of MsLAC1 both in vitro and in vivo. Results Using a comprehensive suite of molecular, biochemical and histochemical analyses, we show that MsLAC1 localizes to cell walls and identify Miscanthus transcription factors capable of regulating MsLAC1 expression. In addition, MsLAC1 complements the Arabidopsis lac4–2 lac17 mutant and recombinant MsLAC1 is able to oxidize monolignol in vitro. Transgenic Arabidopsis plants over-expressing MsLAC1 show higher G-lignin content, although recombinant MsLAC1 seemed to prefer sinapyl alcohol as substrate. Conclusions In summary, our results suggest that MsLAC1 is regulated by secondary cell wall MYB transcription factors and is involved in lignification of xylem fibers. This report identifies MsLAC1 as a promising breeding target in Miscanthus for biofuel and biomaterial applications.

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