Two dynamin-like proteins stabilize FtsZ rings during Streptomyces sporulation
Edited by Susan S. Golden, University of California, San Diego, La Jolla, CA, and approved June 13, 2017 (received for review March 21, 2017)
Significance
Bacterial dynamins were discovered ∼10 y ago and the explosion in genome sequencing has shown that they radiate throughout the bacteria, being present in >1,000 species. In eukaryotes, dynamins play critical roles in the detachment of endocytic vesicles from the plasma membrane, the division of chloroplasts and peroxisomes, and both the fusion and fission of mitochondria. However, in evolutionary terms, dynamins are of bacterial origin, and yet the biological functions of bacterial dynamins remain poorly understood. Here we demonstrate a critical role for dynamins in bacterial cytokinesis, reminiscent of the essential role of eukaryotic dynamins in the division of chloroplasts and mitochondria.
Abstract
During sporulation, the filamentous bacteria Streptomyces undergo a massive cell division event in which the synthesis of ladders of sporulation septa convert multigenomic hyphae into chains of unigenomic spores. This process requires cytokinetic Z-rings formed by the bacterial tubulin homolog FtsZ, and the stabilization of the newly formed Z-rings is crucial for completion of septum synthesis. Here we show that two dynamin-like proteins, DynA and DynB, play critical roles in this process. Dynamins are a family of large, multidomain GTPases involved in key cellular processes in eukaryotes, including vesicle trafficking and organelle division. Many bacterial genomes encode dynamin-like proteins, but the biological function of these proteins has remained largely enigmatic. Using a cell biological approach, we show that the two Streptomyces dynamins specifically localize to sporulation septa in an FtsZ-dependent manner. Moreover, dynamin mutants have a cell division defect due to the decreased stability of sporulation-specific Z-rings, as demonstrated by kymographs derived from time-lapse images of FtsZ ladder formation. This defect causes the premature disassembly of individual Z-rings, leading to the frequent abortion of septum synthesis, which in turn results in the production of long spore-like compartments with multiple chromosomes. Two-hybrid analysis revealed that the dynamins are part of the cell division machinery and that they mediate their effects on Z-ring stability during developmentally controlled cell division via a network of protein–protein interactions involving DynA, DynB, FtsZ, SepF, SepF2, and the FtsZ-positioning protein SsgB.
Data Availability
Data deposition: The data reported in this paper have been deposited in the ArrayExpress database (accession no. E-MTAB-5853).
Acknowledgments
We thank Joe McCormick for the gift of the ftsZ null mutant and the anti-FtsZ antiserum, Matt Bush for sharing bacterial two-hybrid plasmids, Sara Simonini for advice and materials for yeast two-hybrid analysis, Georgia Squyres for helpful discussions, and Grant Calder for technical assistance with the microscope. This work was supported by a Leopoldina Postdoctoral Fellowship (to S.S.), Biotechnology and Biological Sciences Research Council (BBSRC) Grants BB/P001041/1 (to S.S. and M. J. Buttner) and BB/L019825/1 (to M. J. Buttner), BBSRC Institute Strategic Programme Grant BB/J004561/1 to the John Innes Centre, and by grants from the Swedish Research Council (2010-4463 and 2015-05452) and the Crafoord foundation (to K.F.).
Supporting Information
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Movie S1.
Fluorescence time-lapse movie of FtsZ-YPet dynamics during sporulation-specific cell division in the WT (SS12). See also Fig. 3A. (Scale bar: 5 μm.)
Movie S2.
Fluorescence time-lapse movie of FtsZ-YPet dynamics during sporulation-specific cell division in the dynamin mutant (SS14). See also Fig. 3B. (Scale bar: 5 μm.)
References
1
SM Ferguson, P De Camilli, Dynamin, a membrane-remodelling GTPase. Nat Rev Mol Cell Biol 13, 75–88 (2012).
2
HH Low, C Sachse, LA Amos, J Löwe, Structure of a bacterial dynamin-like protein lipid tube provides a mechanism for assembly and membrane curving. Cell 139, 1342–1352 (2009).
3
HH Low, J Löwe, A bacterial dynamin-like protein. Nature 444, 766–769 (2006).
4
M Bramkamp, Structure and function of bacterial dynamin-like proteins. Biol Chem 393, 1203–1214 (2012).
5
O Bohuszewicz, J Liu, HH Low, Membrane remodelling in bacteria. J Struct Biol 196, 3–14 (2016).
6
F Bürmann, N Ebert, S van Baarle, M Bramkamp, A bacterial dynamin-like protein mediating nucleotide-independent membrane fusion. Mol Microbiol 79, 1294–1304 (2011).
7
KA Michie, A Boysen, HH Low, J Møller-Jensen, J Löwe, LeoA, B and C from enterotoxigenic Escherichia coli (ETEC) are bacterial dynamins. PLoS One 9, e107211 (2014).
8
S Ozaki, et al., A replicase clamp-binding dynamin-like protein promotes colocalization of nascent DNA strands and equipartitioning of chromosomes in E. coli. Cell Rep 4, 985–995 (2013).
9
P Sawant, K Eissenberger, L Karier, T Mascher, M Bramkamp, A dynamin-like protein involved in bacterial cell membrane surveillance under environmental stress. Environ Microbiol 18, 2705–2720 (2016).
10
R Colangeli, et al., The Mycobacterium tuberculosis iniA gene is essential for activity of an efflux pump that confers drug tolerance to both isoniazid and ethambutol. Mol Microbiol 55, 1829–1840 (2005).
11
MJ Bush, N Tschowri, S Schlimpert, K Flärdh, MJ Buttner, c-di-GMP signalling and the regulation of developmental transitions in streptomycetes. Nat Rev Microbiol 13, 749–760 (2015).
12
EF Bi, J Lutkenhaus, FtsZ ring structure associated with division in Escherichia coli. Nature 354, 161–164 (1991).
13
J Löwe, LA Amos, Crystal structure of the bacterial cell-division protein FtsZ. Nature 391, 203–206 (1998).
14
DP Haeusser, W Margolin, Splitsville: Structural and functional insights into the dynamic bacterial Z ring. Nat Rev Microbiol 14, 305–319 (2016).
15
J Xiao, ED Goley, Redefining the roles of the FtsZ-ring in bacterial cytokinesis. Curr Opin Microbiol 34, 90–96 (2016).
16
J Willemse, JW Borst, E de Waal, T Bisseling, GP van Wezel, Positive control of cell division: FtsZ is recruited by SsgB during sporulation of Streptomyces. Genes Dev 25, 89–99 (2011).
17
LJ Wu, J Errington, Coordination of cell division and chromosome segregation by a nucleoid occlusion protein in Bacillus subtilis. Cell 117, 915–925 (2004).
18
TG Bernhardt, PA de Boer, SlmA, a nucleoid-associated, FtsZ binding protein required for blocking septal ring assembly over chromosomes in E. coli. Mol Cell 18, 555–564 (2005).
19
PA de Boer, RE Crossley, LI Rothfield, A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in E. coli. Cell 56, 641–649 (1989).
20
R Duman, et al., Structural and genetic analyses reveal the protein SepF as a new membrane anchor for the Z ring. Proc Natl Acad Sci USA 110, E4601–E4610 (2013).
21
P Szwedziak, Q Wang, SM Freund, J Löwe, FtsA forms actin-like protofilaments. EMBO J 31, 2249–2260 (2012).
22
CA Hale, PA de Boer, Direct binding of FtsZ to ZipA, an essential component of the septal ring structure that mediates cell division in E. coli. Cell 88, 175–185 (1997).
23
S Gola, T Munder, S Casonato, R Manganelli, M Vicente, The essential role of SepF in mycobacterial division. Mol Microbiol 97, 560–576 (2015).
24
S Gupta, et al., Essential protein SepF of mycobacteria interacts with FtsZ and MurG to regulate cell growth and division. Microbiology 161, 1627–1638 (2015).
25
CA Hale, et al., Identification of Escherichia coli ZapC (YcbW) as a component of the division apparatus that binds and bundles FtsZ polymers. J Bacteriol 193, 1393–1404 (2011).
26
J Durand-Heredia, E Rivkin, G Fan, J Morales, A Janakiraman, Identification of ZapD as a cell division factor that promotes the assembly of FtsZ in Escherichia coli. J Bacteriol 194, 3189–3198 (2012).
27
JM Durand-Heredia, HH Yu, S De Carlo, CF Lesser, A Janakiraman, Identification and characterization of ZapC, a stabilizer of the FtsZ ring in Escherichia coli. J Bacteriol 193, 1405–1413 (2011).
28
N Grantcharova, U Lustig, K Flärdh, Dynamics of FtsZ assembly during sporulation in Streptomyces coelicolor A3(2). J Bacteriol 187, 3227–3237 (2005).
29
S Schlimpert, K Flärdh, MJ Buttner, Fluorescence time-lapse imaging of the complete S. venezuelae life cycle using a microfluidic device. J Vis Exp, pp. 53863 (2016).
30
K Flärdh, KC Findlay, KF Chater, Association of early sporulation genes with suggested developmental decision points in Streptomyces coelicolor A3(2). Microbiology 145, 2229–2243 (1999).
31
JR McCormick, EP Su, A Driks, R Losick, Growth and viability of Streptomyces coelicolor mutant for the cell division gene ftsZ. Mol Microbiol 14, 243–254 (1994).
32
LW Hamoen, JC Meile, W de Jong, P Noirot, J Errington, SepF, a novel FtsZ-interacting protein required for a late step in cell division. Mol Microbiol 59, 989–999 (2006).
33
ME Gündoğdu, et al., Large ring polymers align FtsZ polymers for normal septum formation. EMBO J 30, 617–626 (2011).
34
CA Konopka, JB Schleede, AR Skop, SY Bednarek, Dynamin and cytokinesis. Traffic 7, 239–247 (2006).
35
SY Miyagishima, H Kuwayama, H Urushihara, H Nakanishi, Evolutionary linkage between eukaryotic cytokinesis and chloroplast division by dynamin proteins. Proc Natl Acad Sci USA 105, 15202–15207 (2008).
36
X Gu, DP Verma, Phragmoplastin, a dynamin-like protein associated with cell plate formation in plants. EMBO J 15, 695–704 (1996).
37
BH Kang, JS Busse, SY Bednarek, Members of the Arabidopsis dynamin-like gene family, ADL1, are essential for plant cytokinesis and polarized cell growth. Plant Cell 15, 899–913 (2003).
38
SY Miyagishima, et al., A plant-specific dynamin-related protein forms a ring at the chloroplast division site. Plant Cell 15, 655–665 (2003).
39
B Feng, H Schwarz, S Jesuthasan, Furrow-specific endocytosis during cytokinesis of zebrafish blastomeres. Exp Cell Res 279, 14–20 (2002).
40
HM Thompson, AR Skop, U Euteneuer, BJ Meyer, MA McNiven, The large GTPase dynamin associates with the spindle midzone and is required for cytokinesis. Curr Biol 12, 2111–2117 (2002).
41
T Kieser, MJ Bibb, MJ Buttner, KF Chater, DA Hopwood Practical Streptomyces Genetics (John Innes Foundation, Norwich, UK, 2000).
42
MJ Bush, MJ Bibb, G Chandra, KC Findlay, MJ Buttner, Genes required for aerial growth, cell division, and chromosome segregation are targets of WhiA before sporulation in Streptomyces venezuelae. MBio 4, e00684-e13 (2013).
43
B Gust, GL Challis, K Fowler, T Kieser, KF Chater, PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proc Natl Acad Sci USA 100, 1541–1546 (2003).
44
B Gust, et al., Lambda red-mediated genetic manipulation of antibiotic-producing Streptomyces. Adv Appl Microbiol 54, 107–128 (2004).
45
LT Fernández-Martínez, MJ Bibb, Use of the meganuclease I-SceI of Saccharomyces cerevisiae to select for gene deletions in actinomycetes. Sci Rep 4, 7100 (2014).
46
J Schwedock, JR McCormick, ER Angert, JR Nodwell, R Losick, Assembly of the cell division protein FtsZ into ladder-like structures in the aerial hyphae of Streptomyces coelicolor. Mol Microbiol 25, 847–858 (1997).
47
KL Griffith, Jr RE Wolf, Systematic mutagenesis of the DNA binding sites for SoxS in the Escherichia coli zwf and fpr promoters: Identifying nucleotides required for DNA binding and transcription activation. Mol Microbiol 40, 1141–1154 (2001).
48
P Slavny, R Little, P Salinas, TA Clarke, R Dixon, Quaternary structure changes in a second Per-Arnt-Sim domain mediate intramolecular redox signal relay in the NifL regulatory protein. Mol Microbiol 75, 61–75 (2010).
49
M Egea-Cortines, H Saedler, H Sommer, Ternary complex formation between the MADS-box proteins SQUAMOSA, DEFICIENS and GLOBOSA is involved in the control of floral architecture in Antirrhinum majus. EMBO J 18, 5370–5379 (1999).
50
MJ Bush, G Chandra, MJ Bibb, KC Findlay, MJ Buttner, Genome-wide chromatin immunoprecipitation sequencing analysis shows that WhiB is a transcription factor that cocontrols its regulon with WhiA to initiate developmental cell division in Streptomyces. MBio 7, e00523-e16 (2016).
51
A Ducret, EM Quardokus, YV Brun, MicrobeJ, a tool for high throughput bacterial cell detection and quantitative analysis. Nat Microbiol 1, 16077 (2016).
52
MJ Bibb, A Domonkos, G Chandra, MJ Buttner, Expression of the chaplin and rodlin hydrophobic sheath proteins in Streptomyces venezuelae is controlled by σ(BldN) and a cognate anti-sigma factor, RsbN. Mol Microbiol 84, 1033–1049 (2012).
53
MS Paget, L Chamberlin, A Atrih, SJ Foster, MJ Buttner, Evidence that the extracytoplasmic function sigma factor sigmaE is required for normal cell wall structure in Streptomyces coelicolor A3(2). J Bacteriol 181, 204–211 (1999).
54
KA Datsenko, BL Wanner, One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97, 6640–6645 (2000).
55
G Karimova, J Pidoux, A Ullmann, D Ladant, A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci USA 95, 5752–5756 (1998).
Information & Authors
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Freely available online through the PNAS open access option.
Data Availability
Data deposition: The data reported in this paper have been deposited in the ArrayExpress database (accession no. E-MTAB-5853).
Submission history
Published online: July 7, 2017
Published in issue: July 25, 2017
Keywords
Acknowledgments
We thank Joe McCormick for the gift of the ftsZ null mutant and the anti-FtsZ antiserum, Matt Bush for sharing bacterial two-hybrid plasmids, Sara Simonini for advice and materials for yeast two-hybrid analysis, Georgia Squyres for helpful discussions, and Grant Calder for technical assistance with the microscope. This work was supported by a Leopoldina Postdoctoral Fellowship (to S.S.), Biotechnology and Biological Sciences Research Council (BBSRC) Grants BB/P001041/1 (to S.S. and M. J. Buttner) and BB/L019825/1 (to M. J. Buttner), BBSRC Institute Strategic Programme Grant BB/J004561/1 to the John Innes Centre, and by grants from the Swedish Research Council (2010-4463 and 2015-05452) and the Crafoord foundation (to K.F.).
Notes
This article is a PNAS Direct Submission.
Authors
Competing Interests
The authors declare no conflict of interest.
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