Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution
Abstract
Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsD in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 angstrom formore »
- Authors:
- Publication Date:
- Research Org.:
- Pennsylvania State Univ., University Park, PA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1337597
- Alternate Identifier(s):
- OSTI ID: 1282049
- Grant/Contract Number:
- SC0001090
- Resource Type:
- Published Article
- Journal Name:
- PLoS ONE
- Additional Journal Information:
- Journal Name: PLoS ONE Journal Volume: 11 Journal Issue: 5; Journal ID: ISSN 1932-6203
- Publisher:
- Public Library of Science (PLoS)
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; particle electron cryomicroscopy; cyclic diguanylic acid; gmp-binding protein; in-vitro synthesis; acetobacter-xylinum; bacterial cellulose; di-gmp; outer-membrane; biosynthesis; subunit
Citation Formats
Du, Juan, Vepachedu, Venkata, Cho, Sung Hyun, Kumar, Manish, Nixon, B. Tracy, and Lai, ed., Hsin-Chih. Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution. United States: N. p., 2016.
Web. doi:10.1371/journal.pone.0155886.
Du, Juan, Vepachedu, Venkata, Cho, Sung Hyun, Kumar, Manish, Nixon, B. Tracy, & Lai, ed., Hsin-Chih. Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution. United States. https://doi.org/10.1371/journal.pone.0155886
Du, Juan, Vepachedu, Venkata, Cho, Sung Hyun, Kumar, Manish, Nixon, B. Tracy, and Lai, ed., Hsin-Chih. Mon .
"Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution". United States. https://doi.org/10.1371/journal.pone.0155886.
@article{osti_1337597,
title = {Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution},
author = {Du, Juan and Vepachedu, Venkata and Cho, Sung Hyun and Kumar, Manish and Nixon, B. Tracy and Lai, ed., Hsin-Chih},
abstractNote = {Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsD in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 angstrom for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. Furthermore, the results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and support the hypothesis that it is the extrusion mechanism and order in linearly arrayed TCs that enables production of crystalline cellulose.},
doi = {10.1371/journal.pone.0155886},
journal = {PLoS ONE},
number = 5,
volume = 11,
place = {United States},
year = {Mon May 23 00:00:00 EDT 2016},
month = {Mon May 23 00:00:00 EDT 2016}
}
https://doi.org/10.1371/journal.pone.0155886
Web of Science
Works referenced in this record:
Genera and species in acetic acid bacteria
journal, June 2008
- Yamada, Yuzo; Yukphan, Pattaraporn
- International Journal of Food Microbiology, Vol. 125, Issue 1
Structural and chemical properties of grass lignocelluloses related to conversion for biofuels
journal, January 2008
- Anderson, William F.; Akin, Danny E.
- Journal of Industrial Microbiology & Biotechnology, Vol. 35, Issue 5
Identification of the uridine 5'-diphosphoglucose (UDP-Glc) binding subunit of cellulose synthase in Acetobacter xylinum using the photoaffinity probe 5-azido-UDP-Glc.
journal, March 1990
- Lin, F. C.; Brown, R. M.; Drake, R. R.
- Journal of Biological Chemistry, Vol. 265, Issue 9
Improved production of bacterial cellulose and its application potential
journal, January 1998
- Vandamme, E. J.; De Baets, S.; Vanbaelen, A.
- Polymer Degradation and Stability, Vol. 59, Issue 1-3
Mechanism of the film thickness increasing during the bacterial production of cellulose on non-agitaded liquid media
journal, November 1995
- Borzani, Walter; de Souza, Silvio J.
- Biotechnology Letters, Vol. 17, Issue 11
Cell-wall carbohydrates and their modification as a resource for biofuels
journal, May 2008
- Pauly, Markus; Keegstra, Kenneth
- The Plant Journal, Vol. 54, Issue 4, p. 559-568
An unusual guanyl oligonucleotide regulates cellulose synthesis in Acetobacter xylinum
journal, July 1985
- Ross, Peter; Aloni, Yehoshua; Weinhouse, Chaim
- FEBS Letters, Vol. 186, Issue 2
The 3D structure of a periplasm-spanning platform required for assembly of group 1 capsular polysaccharides in Escherichia coli
journal, February 2007
- Collins, R. F.; Beis, K.; Dong, C.
- Proceedings of the National Academy of Sciences, Vol. 104, Issue 7
Crystallographic snapshot of cellulose synthesis and membrane translocation
journal, December 2012
- Morgan, Jacob L. W.; Strumillo, Joanna; Zimmer, Jochen
- Nature, Vol. 493, Issue 7431
Three-dimensional structure of human γ-secretase
journal, June 2014
- Lu, Peilong; Bai, Xiao-chen; Ma, Dan
- Nature, Vol. 512, Issue 7513
Recent advances in bacterial cellulose production
journal, February 2005
- Shoda, Makoto; Sugano, Yasushi
- Biotechnology and Bioprocess Engineering, Vol. 10, Issue 1
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
journal, August 1970
- Laemmli, U. K.
- Nature, Vol. 227, Issue 5259, p. 680-685
Accurate determination of local defocus and specimen tilt in electron microscopy
journal, June 2003
- Mindell, Joseph A.; Grigorieff, Nikolaus
- Journal of Structural Biology, Vol. 142, Issue 3
Mechanism of activation of bacterial cellulose synthase by cyclic di-GMP
journal, April 2014
- Morgan, Jacob L. W.; McNamara, Joshua T.; Zimmer, Jochen
- Nature Structural & Molecular Biology, Vol. 21, Issue 5
Pivotal Roles of the Outer Membrane Polysaccharide Export and Polysaccharide Copolymerase Protein Families in Export of Extracellular Polysaccharides in Gram-Negative Bacteria
journal, March 2009
- Cuthbertson, L.; Mainprize, I. L.; Naismith, J. H.
- Microbiology and Molecular Biology Reviews, Vol. 73, Issue 1
Structural Characterization of the Yersinia pestis Type III Secretion System Needle Protein YscF in Complex with Its Heterodimeric Chaperone YscE/YscG
journal, March 2008
- Sun, Ping; Tropea, Joseph E.; Austin, Brian P.
- Journal of Molecular Biology, Vol. 377, Issue 3
The superhelical TPR-repeat domain of O-linked GlcNAc transferase exhibits structural similarities to importin α
journal, September 2004
- Jínek, Martin; Rehwinkel, Jan; Lazarus, Brooke D.
- Nature Structural & Molecular Biology, Vol. 11, Issue 10
Structure of bacterial cellulose synthase subunit D octamer with four inner passageways
journal, October 2010
- Hu, S. -Q.; Gao, Y. -G.; Tajima, K.
- Proceedings of the National Academy of Sciences, Vol. 107, Issue 42
Formation, derivatization and applications of bacterial cellulose
journal, December 1994
- Geyer, U.; Heinze, Th.; Stein, A.
- International Journal of Biological Macromolecules, Vol. 16, Issue 6
Genetic organization of the cellulose synthase operon in Acetobacter xylinum.
journal, October 1990
- Wong, H. C.; Fear, A. L.; Calhoon, R. D.
- Proceedings of the National Academy of Sciences, Vol. 87, Issue 20
Atomic structure of the 26S proteasome lid reveals the mechanism of deubiquitinase inhibition
journal, January 2016
- Dambacher, Corey M.; Worden, Evan J.; Herzik, Mark A.
- eLife, Vol. 5
Effects of CMC Addition on Bacterial Cellulose Production in a Biofilm Reactor and Its Paper Sheets Analysis
journal, March 2011
- Cheng, Kuan-Chen; Catchmark, Jeffrey M.; Demirci, Ali
- Biomacromolecules, Vol. 12, Issue 3
Structure of Full-Length Human PDGFRβ Bound to Its Activating Ligand PDGF-B as Determined by Negative-Stain Electron Microscopy
journal, December 2015
- Chen, Po-Han; Unger, Vinzenz; He, Xiaolin
- Journal of Molecular Biology, Vol. 427, Issue 24
Quantitative analysis of cryo-EM density map segmentation by watershed and scale-space filtering, and fitting of structures by alignment to regions
journal, June 2010
- Pintilie, Grigore D.; Zhang, Junjie; Goddard, Thomas D.
- Journal of Structural Biology, Vol. 170, Issue 3
Localization of c-di-GMP-Binding Protein with the Linear Terminal Complexes of Acetobacter xylinum
journal, October 2001
- Kimura, S.; Chen, H. P.; Saxena, I. M.
- Journal of Bacteriology, Vol. 183, Issue 19
A new gene required for cellulose production and a gene encoding cellulolytic activity in Acetobacter xylinum are colocalized with the bcs operon.
journal, February 1994
- Standal, R.; Iversen, T. G.; Coucheron, D. H.
- Journal of Bacteriology, Vol. 176, Issue 3
Processing of cellulose synthase (AcsAB) from Gluconacetobacter hansenii 23769
journal, January 2013
- Iyer, Prashanti R.; Liu, Yu-An; Deng, Ying
- Archives of Biochemistry and Biophysics, Vol. 529, Issue 2
Biosynthesis of Callose and Cellulose by Detergent Extracts of Tobacco Cell Membranes and Quantification of the Polymers Synthesized in vitro
journal, February 2010
- Cifuentes, Carolina; Bulone, Vincent; Emons, Anne Mie C.
- Journal of Integrative Plant Biology, Vol. 52, Issue 2
In vitro synthesis of cellulose II from a cytoplasmic membrane fraction of Acetobacter xylinum
journal, October 1987
- Bureau, T. E.; Brown, R. M.
- Proceedings of the National Academy of Sciences, Vol. 84, Issue 20
Atomic model of the F420-reducing [NiFe] hydrogenase by electron cryo-microscopy using a direct electron detector
journal, February 2014
- Allegretti, Matteo; Mills, Deryck J.; McMullan, Greg
- eLife, Vol. 3
Description of Komagataeibacter gen. nov., with proposals of new combinations (Acetobacteraceae)
journal, January 2012
- Yamada, Yuzo; Yukphan, Pattaraporn; Lan Vu, Huong Thi
- The Journal of General and Applied Microbiology, Vol. 58, Issue 5
AcsA–AcsB: The core of the cellulose synthase complex from Gluconacetobacter hansenii ATCC23769
journal, January 2016
- McManus, John B.; Deng, Ying; Nagachar, Nivedita
- Enzyme and Microbial Technology, Vol. 82
Genome Sequence of a Cellulose-Producing Bacterium, Gluconacetobacter hansenii ATCC 23769
journal, June 2010
- Iyer, P. R.; Geib, S. M.; Catchmark, J.
- Journal of Bacteriology, Vol. 192, Issue 16
Cellulose complementing factor (Ccp) is a new member of the cellulose synthase complex (terminal complex) in Acetobacter xylinum
journal, June 2013
- Sunagawa, Naoki; Fujiwara, Takaaki; Yoda, Takanori
- Journal of Bioscience and Bioengineering, Vol. 115, Issue 6
Optimal Determination of Particle Orientation, Absolute Hand, and Contrast Loss in Single-particle Electron Cryomicroscopy
journal, October 2003
- Rosenthal, Peter B.; Henderson, Richard
- Journal of Molecular Biology, Vol. 333, Issue 4
Regulation of cellulose synthesis in Acetobacter xylinum by cyclic diguanylic acid
journal, January 1987
- Ross, P.; Weinhouse, H.; Aloni, Y.
- Nature, Vol. 325, Issue 6101
Nanocelluloses: A New Family of Nature-Based Materials
journal, May 2011
- Klemm, Dieter; Kramer, Friederike; Moritz, Sebastian
- Angewandte Chemie International Edition, Vol. 50, Issue 24
EMAN2: An extensible image processing suite for electron microscopy
journal, January 2007
- Tang, Guang; Peng, Liwei; Baldwin, Philip R.
- Journal of Structural Biology, Vol. 157, Issue 1
Characterization of genes in the cellulose-synthesizing operon (acs operon) of Acetobacter xylinum: implications for cellulose crystallization.
journal, September 1994
- Saxena, I. M.; Kudlicka, K.; Okuda, K.
- Journal of Bacteriology, Vol. 176, Issue 18
Structural flexibility of the G s -helical domain in the 2-adrenoceptor Gs complex
journal, September 2011
- Westfield, G. H.; Rasmussen, S. G. F.; Su, M.
- Proceedings of the National Academy of Sciences, Vol. 108, Issue 38
Structural basis for alginate secretion across the bacterial outer membrane
journal, July 2011
- Whitney, John C.; Hay, Iain D.; Li, Canhui
- Proceedings of the National Academy of Sciences, Vol. 108, Issue 32
Visualizing density maps with UCSF Chimera
journal, January 2007
- Goddard, Thomas D.; Huang, Conrad C.; Ferrin, Thomas E.
- Journal of Structural Biology, Vol. 157, Issue 1
Cloning and sequencing of the cellulose synthase catalytic subunit gene of Acetobacter xylinum
journal, November 1990
- Saxena, Inder M.; Lin, Fong Chyr; Brown, R. Malcolm
- Plant Molecular Biology, Vol. 15, Issue 5
Iterative Stable Alignment and Clustering of 2D Transmission Electron Microscope Images
journal, February 2012
- Yang, Zhengfan; Fang, Jia; Chittuluru, Johnathan
- Structure, Vol. 20, Issue 2
Cellulose structure and biosynthesis: What is in store for the 21st century?
journal, January 2003
- Brown, R. Malcolm
- Journal of Polymer Science Part A: Polymer Chemistry, Vol. 42, Issue 3
Visualization of arrestin recruitment by a G-protein-coupled receptor
journal, June 2014
- Shukla, Arun K.; Westfield, Gerwin H.; Xiao, Kunhong
- Nature, Vol. 512, Issue 7513
Visualization of pores (export sites) correlated with cellulose production in the envelope of the gram-negative bacterium Acetobacter xylinum
journal, March 1979
- Zaar, K.
- The Journal of Cell Biology, Vol. 80, Issue 3
Structure of the heterotrimeric complex that regulates type III secretion needle formation
journal, April 2007
- Quinaud, M.; Ple, S.; Job, V.
- Proceedings of the National Academy of Sciences, Vol. 104, Issue 19
AlgK Is a TPR-Containing Protein and the Periplasmic Component of a Novel Exopolysaccharide Secretin
journal, February 2010
- Keiski, Carrie-Lynn; Harwich, Michael; Jain, Sumita
- Structure, Vol. 18, Issue 2
Structural and functional characteristics of bacterial biofilms
journal, August 2010
- Smirnova, T. A.; Didenko, L. V.; Azizbekyan, R. R.
- Microbiology, Vol. 79, Issue 4
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
journal, May 1976
- Bradford, Marion M.
- Analytical Biochemistry, Vol. 72, Issue 1-2
Structure of the L Protein of Vesicular Stomatitis Virus from Electron Cryomicroscopy
journal, July 2015
- Liang, Bo; Li, Zongli; Jenni, Simon
- Cell, Vol. 162, Issue 2
The cyclic diguanylic acid regulatory system of cellulose synthesis in Acetobacter xylinum. Chemical synthesis and biological activity of cyclic nucleotide dimer, trimer, and phosphothioate derivatives.
journal, November 1990
- Ross, P.; Mayer, R.; Weinhouse, H.
- Journal of Biological Chemistry, Vol. 265, Issue 31
Enhanced production of bacterial cellulose by using a biofilm reactor and its material property analysis
journal, January 2009
- Cheng, Kuan-Chen; Catchmark, Jeff M.; Demirci, Ali
- Journal of Biological Engineering, Vol. 3, Issue 1
BcsA and BcsB form the catalytically active core of bacterial cellulose synthase sufficient for in vitro cellulose synthesis
journal, October 2013
- Omadjela, O.; Narahari, A.; Strumillo, J.
- Proceedings of the National Academy of Sciences, Vol. 110, Issue 44
Identification and Characterization of Non-Cellulose-Producing Mutants of Gluconacetobacter hansenii Generated by Tn5 Transposon Mutagenesis
journal, September 2013
- Deng, Y.; Nagachar, N.; Xiao, C.
- Journal of Bacteriology, Vol. 195, Issue 22
TRPV1 structures in distinct conformations reveal activation mechanisms
journal, December 2013
- Cao, Erhu; Liao, Maofu; Cheng, Yifan
- Nature, Vol. 504, Issue 7478
Identification of a new gene in an operon for cellulose biosynthesis in Acetobacter xylinum
journal, June 1991
- Saxena, Inder M.; Lin, Fong Chyr; Brown, R. Malcolm
- Plant Molecular Biology, Vol. 16, Issue 6
Structure of the TRPV1 ion channel determined by electron cryo-microscopy
journal, December 2013
- Liao, Maofu; Cao, Erhu; Julius, David
- Nature, Vol. 504, Issue 7478
Antimicrobial Bacterial Cellulose-Silver Nanoparticles Composite Membranes
journal, January 2011
- Barud, Hernane S.; Regiani, Thaís; Marques, Rodrigo F. C.
- Journal of Nanomaterials, Vol. 2011
Bacterial cellulose biosynthesis: diversity of operons, subunits, products, and functions
journal, September 2015
- Römling, Ute; Galperin, Michael Y.
- Trends in Microbiology, Vol. 23, Issue 9
Structure of the Sec61 channel opened by a signal sequence
journal, December 2015
- Voorhees, R. M.; Hegde, R. S.
- Science, Vol. 351, Issue 6268
Solubilization of the UDP-glucose:1,4-beta-D-glucan 4-beta-D-glucosyltransferase (cellulose synthase) from Acetobacter xylinum. A comparison of regulatory properties with those of the membrane-bound form of the enzyme.
journal, April 1983
- Aloni, Y.; Cohen, R.; Benziman, M.
- Journal of Biological Chemistry, Vol. 258, Issue 7
c-di-GMP-binding protein, a new factor regulating cellulose synthesis in Acetobacter xylinum
journal, October 1997
- Weinhouse, Haim; Sapir, Shai; Amikam, Dorit
- FEBS Letters, Vol. 416, Issue 2
Cryo-EM structure of the Plasmodium falciparum 80S ribosome bound to the anti-protozoan drug emetine
journal, June 2014
- Wong, Wilson; Bai, Xiao-chen; Brown, Alan
- eLife, Vol. 3
In vitro synthesis of cellulose microfibrils by a membrane protein from protoplasts of the non-vascular plant Physcomitrella patens
journal, August 2015
- Cho, Sung Hyun; Du, Juan; Sines, Ian
- Biochemical Journal, Vol. 470, Issue 2
Tilt-Pair Analysis of Images from a Range of Different Specimens in Single-Particle Electron Cryomicroscopy
journal, November 2011
- Henderson, Richard; Chen, Shaoxia; Chen, James Z.
- Journal of Molecular Biology, Vol. 413, Issue 5
Cellulose biosynthesis and function in bacteria.
journal, January 1991
- Ross, P.; Mayer, R.; Benziman, M.
- Microbiological Reviews, Vol. 55, Issue 1
Structure of the Yeast Mitochondrial Large Ribosomal Subunit
journal, March 2014
- Amunts, A.; Brown, A.; Bai, X. -c.
- Science, Vol. 343, Issue 6178
Subdivision of the genus Gluconacetobacter Yamada, Hoshino and Ishikawa 1998: the proposal of Komagatabacter gen. nov., for strains accommodated to the Gluconacetobacter xylinus group in the α-Proteobacteria
journal, June 2011
- Yamada, Yuzo; Yukphan, Pattaraporn; Vu, Huong Thi Lan
- Annals of Microbiology, Vol. 62, Issue 2