Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum

Shin, Jae Yen; Lopez-Garrido, Javier; Lee, Sang-Hyuk; Diaz-Celis, Cesar; Fleming, Tinya; Bustamante, Carlos; Pogliano, Kit

doi:10.7554/eLife.06474

Title: Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum

Journal Article · Thu May 07 00:00:00 EDT 2015 · eLife

DOI:https://doi.org/10.7554/eLife.06474· OSTI ID:1180127

Shin, Jae Yen ^[1]; Lopez-Garrido, Javier ^[2]; Lee, Sang-Hyuk ^[1]; Diaz-Celis, Cesar ^[1]; Fleming, Tinya ^[2]; Bustamante, Carlos ^[3]; Pogliano, Kit ^[2]

Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States
Division of Biological Sciences, University of California, San Diego, La Jolla, United States
Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States, Jason L Choy Laboratory of Single Molecule Biophysics, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States, QB3 Institute, University of California, Berkeley, Berkeley, United States, Kavli Energy NanoSciences Institute, University of California, Berkeley, Berkeley, United States

SpoIIIE is a membrane-anchored DNA translocase that localizes to the septal midpoint to mediate chromosome translocation and membrane fission during Bacillus subtilis sporulation. Here we use cell-specific protein degradation and quantitative photoactivated localization microscopy in strains with a thick sporulation septum to investigate the architecture and function of the SpoIIIE DNA translocation complex in vivo. We were able to visualize SpoIIIE complexes with approximately equal numbers of molecules in the mother cell and the forespore. Cell-specific protein degradation showed that only the mother cell complex is required to translocate DNA into the forespore, whereas degradation in either cell reverses membrane fission. Our data suggest that SpoIIIE assembles a coaxially paired channel for each chromosome arm comprised of one hexamer in each cell to maintain membrane fission during DNA translocation. We show that SpoIIIE can operate, in principle, as a bi-directional motor that exports DNA.

View Journal Article

Cite

Export

Save

Research Organization:: Univ. of California, Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences and Engineering Division; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1180127

Alternate ID(s):: OSTI ID: 1184781; OSTI ID: 1197950

Journal Information:: eLife, Journal Name: eLife Vol. 4; ISSN 2050-084X

Publisher:: eLife Sciences Publications, Ltd.Copyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 29 works

Citation information provided by
Web of Science

References (47)

Molecular Mechanism of Sequence-Directed DNA Loading and Translocation by FtsK Löwe, Jan; Ellonen, Antti; Allen, Mark D. Molecular Cell, Vol. 31, Issue 4 https://doi.org/10.1016/j.molcel.2008.05.027	journal	August 2008
spoIIQ , a forespore‐expressed gene required for engulfment in Bacillus subtilis Londoño‐Vallejo, J. ‐Arturo; Fréhel, Claude; Stragier, Patrick Molecular Microbiology, Vol. 24, Issue 1 https://doi.org/10.1046/j.1365-2958.1997.3181680.x	journal	April 1997
Commitment to sporulation in Bacillus subtilis and its relationship to development of actinomycin resistance Sterlini, Janet M.; Mandelstam, J. Biochemical Journal, Vol. 113, Issue 1 https://doi.org/10.1042/bj1130029	journal	June 1969
Engulfment during sporulation in Bacillus subtilis is governed by a multi-protein complex containing tandemly acting autolysins: Multi-protein complex containing autolysins Chastanet, Arnaud; Losick, Richard Molecular Microbiology, Vol. 64, Issue 1 https://doi.org/10.1111/j.1365-2958.2007.05652.x	journal	March 2007
Inducible protein degradation in Bacillus subtilis using heterologous peptide tags and adaptor proteins to target substrates to the protease ClpXP Griffith, Kevin L.; Grossman, Alan D. Molecular Microbiology https://doi.org/10.1111/j.1365-2958.2008.06467.x	journal	November 2008
Evidence that the SpoIIIE DNA translocase participates in membrane fusion during cytokinesis and engulfment Liu, Nai-Jia Linda; Dutton, Rachel J.; Pogliano, Kit Molecular Microbiology, Vol. 59, Issue 4 https://doi.org/10.1111/j.1365-2958.2005.05004.x	journal	February 2006
TrwB, the coupling protein involved in DNA transport during bacterial conjugation, is a DNA-dependent ATPase Tato, I.; Zunzunegui, S.; de la Cruz, F. Proceedings of the National Academy of Sciences, Vol. 102, Issue 23 https://doi.org/10.1073/pnas.0503402102	journal	May 2005
Structure and DNA-binding properties of the Bacillus subtilis SpoIIIE DNA translocase revealed by single-molecule and electron microscopies Cattoni, Diego I.; Thakur, Shreyasi; Godefroy, Cedric Nucleic Acids Research, Vol. 42, Issue 4 https://doi.org/10.1093/nar/gkt1231	journal	November 2013
Role of Bacillus subtilis SpoIIIE in DNA Transport Across the Mother Cell-Prespore Division Septum Bath, J. Science, Vol. 290, Issue 5493 https://doi.org/10.1126/science.290.5493.995	journal	November 2000
Trans-complex formation by proteolipid channels in the terminal phase of membrane fusion Peters, Christopher; Bayer, Martin J.; Bühler, Susanne Nature, Vol. 409, Issue 6820 https://doi.org/10.1038/35054500	journal	February 2001
Comparative genomics of the FtsK-HerA superfamily of pumping ATPases: implications for the origins of chromosome segregation, cell division and viral capsid packaging Iyer, L. M. Nucleic Acids Research, Vol. 32, Issue 17 https://doi.org/10.1093/nar/gkh828	journal	September 2004
Learning Rates and States from Biophysical Time Series: A Bayesian Approach to Model Selection and Single-Molecule FRET Data Bronson, Jonathan E.; Fei, Jingyi; Hofman, Jake M. Biophysical Journal, Vol. 97, Issue 12 https://doi.org/10.1016/j.bpj.2009.09.031	journal	December 2009
SpoIIIE mechanism of directional translocation involves target search coupled to sequence-dependent motor stimulation Cattoni, Diego I.; Chara, Osvaldo; Godefroy, Cédric EMBO reports, Vol. 14, Issue 5 https://doi.org/10.1038/embor.2013.39	journal	April 2013
Cell-specific SpoIIIE assembly and DNA translocation polarity are dictated by chromosome orientation Becker, Eric C.; Pogliano, Kit Molecular Microbiology, Vol. 66, Issue 5 https://doi.org/10.1111/j.1365-2958.2007.05992.x	journal	December 2007
Counting molecules in single organelles with superresolution microscopy allows tracking of the endosome maturation trajectory Puchner, E. M.; Walter, J. M.; Kasper, R. Proceedings of the National Academy of Sciences, Vol. 110, Issue 40 https://doi.org/10.1073/pnas.1309676110	journal	September 2013
SpoIID-Mediated Peptidoglycan Degradation Is Required throughout Engulfment during Bacillus subtilis Sporulation Gutierrez, J.; Smith, R.; Pogliano, K. Journal of Bacteriology, Vol. 192, Issue 12 https://doi.org/10.1128/JB.00127-10	journal	April 2010
Peptidoglycan transformations during Bacillus subtilis sporulation : Peptidoglycan remodelling Tocheva, Elitza I.; López-Garrido, Javier; Hughes, H. Velocity Molecular Microbiology, Vol. 88, Issue 4 https://doi.org/10.1111/mmi.12201	journal	March 2013
Fusion needs more than SNAREs Almers, Wolfhard Nature, Vol. 409, Issue 6820 https://doi.org/10.1038/35054637	journal	February 2001
An in vivo membrane fusion assay implicates SpoIIIE in the final stages of engulfment during Bacillus subtilis sporulation Sharp, M. D.; Pogliano, K. Proceedings of the National Academy of Sciences, Vol. 96, Issue 25 https://doi.org/10.1073/pnas.96.25.14553	journal	December 1999
Lipid bilayer thickness varies linearly with acyl chain length in fluid phosphatidylcholine vesicles Lewis, Barbara A.; Engelman, Donald M. Journal of Molecular Biology, Vol. 166, Issue 2 https://doi.org/10.1016/S0022-2836(83)80007-2	journal	May 1983
A vital stain for studying membrane dynamics in bacteria: a novel mechanism controlling septation during Bacillus subtilis sporulation Pogliano, Joe; Osborne, Nick; Sharp, Marc D. Molecular Microbiology, Vol. 31, Issue 4 https://doi.org/10.1046/j.1365-2958.1999.01255.x	journal	February 1999
A cytoskeleton-like role for the bacterial cell wall during engulfment of the Bacillus subtilis forespore Abanes-De Mello, A. Genes & Development, Vol. 16, Issue 24 https://doi.org/10.1101/gad.1039902	journal	December 2002
The ATPase SpoIIIE Transports DNA across Fused Septal Membranes during Sporulation in Bacillus subtilis Burton, Briana M.; Marquis, Kathleen A.; Sullivan, Nora L. Cell, Vol. 131, Issue 7 https://doi.org/10.1016/j.cell.2007.11.009	journal	December 2007
Septal localization of the SpoIIIE chromosome partitioning protein in Bacillus subtilis Wu, L. J. The EMBO Journal, Vol. 16, Issue 8 https://doi.org/10.1093/emboj/16.8.2161	journal	April 1997
Genetic regulation of morphogenesis in Bacillus subtilis: roles of sigma E and sigma F in prespore engulfment. Illing, N.; Errington, J. Journal of Bacteriology, Vol. 173, Issue 10 https://doi.org/10.1128/jb.173.10.3159-3169.1991	journal	May 1991
FtsK is an essential cell division protein that is localized to the septum and induced as part of the SOS response Wang, Lilin; Lutkenhaus, Joe Molecular Microbiology, Vol. 29, Issue 3 https://doi.org/10.1046/j.1365-2958.1998.00958.x	journal	August 1998
Sequence-directed DNA export guides chromosome translocation during sporulation in Bacillus subtilis Ptacin, Jerod L.; Nollmann, Marcelo; Becker, Eric C. Nature Structural & Molecular Biology, Vol. 15, Issue 5 https://doi.org/10.1038/nsmb.1412	journal	April 2008
Sequence-Directed DNA Translocation by Purified FtsK Pease, P. J. Science, Vol. 307, Issue 5709 https://doi.org/10.1126/science.1104885	journal	January 2005
Recruitment, Assembly, and Molecular Architecture of the SpoIIIE DNA Pump Revealed by Superresolution Microscopy Fiche, Jean-Bernard; Cattoni, Diego I.; Diekmann, Nele PLoS Biology, Vol. 11, Issue 5 https://doi.org/10.1371/journal.pbio.1001557	journal	May 2013
DNA segregation by the bacterial actin AlfA during Bacillus subtilis growth and development Becker, Eric; Herrera, Nick C.; Gunderson, Felizza Q. The EMBO Journal, Vol. 25, Issue 24 https://doi.org/10.1038/sj.emboj.7601443	journal	November 2006
Double-Stranded DNA Translocation: Structure and Mechanism of Hexameric FtsK Massey, Thomas H.; Mercogliano, Christopher P.; Yates, James Molecular Cell, Vol. 23, Issue 4 https://doi.org/10.1016/j.molcel.2006.06.019	journal	August 2006
Transcriptional control of the Bacillus subtilis spoIID gene. Rong, S.; Rosenkrantz, M. S.; Sonenshein, A. L. Journal of Bacteriology, Vol. 165, Issue 3 https://doi.org/10.1128/jb.165.3.771-779.1986	journal	March 1986
Membrane-associated DNA Transport Machines Burton, B.; Dubnau, D. Cold Spring Harbor Perspectives in Biology, Vol. 2, Issue 7 https://doi.org/10.1101/cshperspect.a000406	journal	June 2010
Plasmids for ectopic integration in Bacillus subtilis Guérout-Fleury, Anne-Marie; Frandsen, Niels; Stragier, Patrick Gene, Vol. 180, Issue 1-2 https://doi.org/10.1016/S0378-1119(96)00404-0	journal	January 1996
DNA transport in bacteria Errington, Jeff; Bath, Jonathan; Wu, Ling Juan Nature Reviews Molecular Cell Biology, Vol. 2, Issue 7 https://doi.org/10.1038/35080005	journal	July 2001
Counting single photoactivatable fluorescent molecules by photoactivated localization microscopy (PALM) Lee, S. -H.; Shin, J. Y.; Lee, A. Proceedings of the National Academy of Sciences, Vol. 109, Issue 43 https://doi.org/10.1073/pnas.1215175109	journal	October 2012
Role of Cell-Specific SpoIIIE Assembly in Polarity of DNA Transfer Sharp, M. D. Science, Vol. 295, Issue 5552 https://doi.org/10.1126/science.1066274	journal	January 2002
Kernel density estimation via diffusion Botev, Z. I.; Grotowski, J. F.; Kroese, D. P. The Annals of Statistics, Vol. 38, Issue 5 https://doi.org/10.1214/10-AOS799	journal	October 2010
Use of lacZ Gene Fusions to Determine the Dependence Pattern of the Sporulation Gene spoIID in spo Mutants of Bacillus subtilis Clarke, S.; Lopez-Diaz, I.; Mandelstam, J. Microbiology, Vol. 132, Issue 11 https://doi.org/10.1099/00221287-132-11-2987	journal	November 1986
Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate Durisic, Nela; Laparra-Cuervo, Lara; Sandoval-Álvarez, Ángel Nature Methods, Vol. 11, Issue 2 https://doi.org/10.1038/nmeth.2784	journal	January 2014
The role of the sporulation gene spollIE in the regulation of prespore-specific gene expression in Bacillus subtilis Foulger, D.; Errington, J. Molecular Microbiology, Vol. 3, Issue 9 https://doi.org/10.1111/j.1365-2958.1989.tb00275.x	journal	September 1989
SpoIIIE Protein Achieves Directional DNA Translocation through Allosteric Regulation of ATPase Activity by an Accessory Domain Besprozvannaya, Marina; Pivorunas, Valerie L.; Feldman, Zachary Journal of Biological Chemistry, Vol. 288, Issue 40 https://doi.org/10.1074/jbc.M113.484055	journal	August 2013
Dynamic SpoIIIE assembly mediates septal membrane fission during Bacillus subtilis sporulation Fleming, T. C.; Shin, J. Y.; Lee, S. H. Genes & Development, Vol. 24, Issue 11 https://doi.org/10.1101/gad.1925210	journal	June 2010
The Membrane Domain of SpoIIIE Is Required for Membrane Fusion during Bacillus subtilis Sporulation Sharp, M. D.; Pogliano, K. Journal of Bacteriology, Vol. 185, Issue 6 https://doi.org/10.1128/JB.185.6.2005-2008.2003	journal	March 2003
Bacillus subtilis spoIIIE protein required for DNA segregation during asymmetric cell division Wu, L.; Errington, J. Science, Vol. 264, Issue 5158 https://doi.org/10.1126/science.8160014	journal	April 1994
Optimization of membrane protein overexpression and purification using GFP fusions Drew, David; Lerch, Mirjam; Kunji, Edmund Nature Methods, Vol. 3, Issue 4 https://doi.org/10.1038/nmeth0406-303	journal	April 2006
Localization of Cell Division Protein FtsK to theEscherichia coli Septum and Identification of a Potential N-Terminal Targeting Domain Yu, Xuan-chuan; Tran, Anthony H.; Sun, Qin Journal of Bacteriology, Vol. 180, Issue 5 https://doi.org/10.1128/JB.180.5.1296-1304.1998	journal	March 1998

Similar Records

A DNA-Binding Protein Tunes Septum Placement during Bacillus subtilis Sporulation

Journal Article · Thu Aug 15 00:00:00 EDT 2019 · Journal of Bacteriology · OSTI ID:1180127

Brown, Emily E.; Miller, Allyssa K.; Krieger, Inna V.; +3 more

Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

Journal Article · Fri Oct 09 00:00:00 EDT 2015 · eLife · OSTI ID:1180127

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

Mechanochemical coupling and bi-phasic force-velocity dependence in the ultra-fast ring ATPase SpoIIIE

Journal Article · Mon Mar 05 00:00:00 EST 2018 · eLife · OSTI ID:1180127

Liu, Ninning; Chistol, Gheorghe; Cui, Yuanbo; +1 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES

Title: Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum

Citation Formats

References (47)

Similar Records

Related Subjects