Coiled-coil protein Scy is a key component of a multiprotein assembly controlling polarized growth in Streptomyces

Polarized growth in eukaryotes requires polar multiprotein complexes. Here, we establish that selection and maintenance of cell polarity for growth also requires a dedicated multiprotein assembly in the filamentous bacterium, Streptomyces coelicolor. We present evidence for a tip organizing center and confirm two of its main components: Scy (Streptomyces cytoskeletal element), a unique bacterial coiled-coil protein with an unusual repeat periodicity, and the known polarity determinant DivIVA. We also establish a link between the tip organizing center and the filament-forming protein FilP. Interestingly, both deletion and overproduction of Scy generated multiple polarity centers, suggesting a mechanism wherein Scy can both promote and limit the number of emerging polarity centers via the organization of the Scy-DivIVA assemblies. We propose that Scy is a molecular “assembler,” which, by sequestering DivIVA, promotes the establishment of new polarity centers for de novo tip formation during branching, as well as supporting polarized growth at existing hyphal tips.

[1]  K. Flärdh,et al.  Dynamics of FtsZ Assembly during Sporulation in Streptomyces coelicolor A3(2) , 2005, Journal of bacteriology.

[2]  A. Lupas,et al.  Predicting coiled coils from protein sequences , 1991, Science.

[3]  I. Meier,et al.  Coiled-coil protein composition of 22 proteomes – differences and common themes in subcellular infrastructure and traffic control , 2005, BMC Evolutionary Biology.

[4]  R. Losick,et al.  Growth and viability of Streptomyces coelicolor mutant for the cell division gene ftsZ , 1994, Molecular microbiology.

[5]  H. Lam,et al.  A Landmark Protein Essential for Establishing and Perpetuating the Polarity of a Bacterial Cell , 2006, Cell.

[6]  W. Wohlleben,et al.  Proteins encoded by the mre gene cluster in Streptomyces coelicolor A3(2) cooperate in spore wall synthesis , 2011, Molecular microbiology.

[7]  W. Margolin,et al.  Sculpting the Bacterial Cell , 2009, Current Biology.

[8]  J. Errington,et al.  Control of Cell Morphogenesis in Bacteria Two Distinct Ways to Make a Rod-Shaped Cell , 2003, Cell.

[9]  Z. Deng,et al.  A Cellulose Synthase-Like Protein Involved in Hyphal Tip Growth and Morphological Differentiation in Streptomyces , 2008, Journal of bacteriology.

[10]  Andrei N. Lupas,et al.  The structure of α-helical coiled coils , 2005 .

[11]  L. Shapiro,et al.  A Polymeric Protein Anchors the Chromosomal Origin/ParB Complex at a Bacterial Cell Pole , 2008, Cell.

[12]  Yong-Gyun Jung,et al.  The Ser/Thr protein kinase AfsK regulates polar growth and hyphal branching in the filamentous bacteria Streptomyces , 2012, Proceedings of the National Academy of Sciences.

[13]  J. Errington,et al.  Dispersed mode of Staphylococcus aureus cell wall synthesis in the absence of the division machinery , 2003, Molecular microbiology.

[14]  K. Flärdh Essential role of DivIVA in polar growth and morphogenesis in Streptomyces coelicolor A3(2) , 2003, Molecular microbiology.

[15]  N. Ausmees,et al.  Intermediate filament-like proteins in bacteria and a cytoskeletal function in Streptomyces , 2008, Molecular microbiology.

[16]  J. Errington,et al.  Nucleoid occlusion and bacterial cell division , 2011, Nature Reviews Microbiology.

[17]  Amy E Keating,et al.  Analysis of coiled-coil interactions between core proteins of the spindle pole body. , 2008, Biochemistry.

[18]  C. Jacobs-Wagner,et al.  The Bacterial Cytoskeleton An Intermediate Filament-Like Function in Cell Shape , 2003, Cell.

[19]  Klas Flärdh,et al.  Streptomyces morphogenetics: dissecting differentiation in a filamentous bacterium , 2009, Nature Reviews Microbiology.

[20]  H. McAdams,et al.  Caulobacter PopZ forms a polar subdomain dictating sequential changes in pole composition and function , 2010, Molecular microbiology.

[21]  W. Wohlleben,et al.  Unique conjugation mechanism in mycelial streptomycetes: a DNA‐binding ATPase translocates unprocessed plasmid DNA at the hyphal tip , 2006, Molecular microbiology.

[22]  E. Huitema,et al.  Bacterial Birth Scar Proteins Mark Future Flagellum Assembly Site , 2006, Cell.

[23]  R. Krämer,et al.  A synthetic Escherichia coli system identifies a conserved origin tethering factor in Actinobacteria , 2012, Molecular microbiology.

[24]  Bruce L. Goode,et al.  The Yeast Actin Cytoskeleton: from Cellular Function to Biochemical Mechanism , 2006, Microbiology and Molecular Biology Reviews.

[25]  J. Walshaw,et al.  A novel coiled-coil repeat variant in a class of bacterial cytoskeletal proteins. , 2010, Journal of structural biology.

[26]  K. Chater,et al.  Alignment of multiple chromosomes along helical ParA scaffolding in sporulating Streptomyces hyphae , 2007, Molecular microbiology.

[27]  A. Keating,et al.  Specific coiled-coil interactions contribute to a global model of the structure of the spindle pole body. , 2010, Journal of structural biology.

[28]  T. Leonard,et al.  Features critical for membrane binding revealed by DivIVA crystal structure , 2010, The EMBO journal.

[29]  Terry K. Smith,et al.  Cardiolipin synthase is required for Streptomyces coelicolor morphogenesis , 2012, Molecular microbiology.

[30]  Gilles P van Wezel,et al.  MreB of Streptomyces coelicolor is not essential for vegetative growth but is required for the integrity of aerial hyphae and spores , 2006, Molecular microbiology.

[31]  Yu-Ling Shih,et al.  The Bacterial Cytoskeleton , 2006, Microbiology and Molecular Biology Reviews.

[32]  Kumaran S Ramamurthi,et al.  Negative membrane curvature as a cue for subcellular localization of a bacterial protein , 2009, Proceedings of the National Academy of Sciences.

[33]  W. Nelson,et al.  Adaptation of core mechanisms to generate cell polarity , 2003, Nature.

[34]  J. Willemse,et al.  Positive control of cell division: FtsZ is recruited by SsgB during sporulation of Streptomyces. , 2011, Genes & development.

[35]  P. Datta,et al.  Novel role of Wag31 in protection of mycobacteria under oxidative stress , 2009, Molecular microbiology.

[36]  S. Cantlay,et al.  Domains involved in the in vivo function and oligomerization of apical growth determinant DivIVA in Streptomyces coelicolor. , 2009, FEMS microbiology letters.

[37]  K. Flärdh,et al.  The MreB-Like Protein Mbl of Streptomyces coelicolor A3(2) Depends on MreB for Proper Localization and Contributes to Spore Wall Synthesis , 2011, Journal of bacteriology.

[38]  David M. Richards,et al.  Mechanistic Basis of Branch-Site Selection in Filamentous Bacteria , 2012, PLoS Comput. Biol..

[39]  A. Lupas,et al.  The structure of alpha-helical coiled coils. , 2005, Advances in protein chemistry.

[40]  C. Jacobs-Wagner,et al.  Bacterial intermediate filaments: in vivo assembly, organization, and dynamics of crescentin. , 2009, Genes & development.

[41]  J. Willemse,et al.  Loss of the controlled localization of growth stage‐specific cell‐wall synthesis pleiotropically affects developmental gene expression in an ssgA mutant of Streptomyces coelicolor , 2007, Molecular microbiology.

[42]  T. Kieser Practical streptomyces genetics , 2000 .

[43]  J. Errington,et al.  Localisation of DivIVA by targeting to negatively curved membranes , 2009, The EMBO journal.

[44]  D. Ladant,et al.  A bacterial two-hybrid system based on a reconstituted signal transduction pathway. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[45]  G. Jensen,et al.  A Self-Associating Protein Critical for Chromosome Attachment, Division, and Polar Organization in Caulobacter , 2008, Cell.

[46]  Antje M. Hempel,et al.  Assemblies of DivIVA Mark Sites for Hyphal Branching and Can Establish New Zones of Cell Wall Growth in Streptomyces coelicolor , 2008, Journal of bacteriology.

[47]  K. Flärdh Cell polarity and the control of apical growth in Streptomyces. , 2010, Current opinion in microbiology.