title: Time-lapse microscopy of Streptomyces coelicolor growth and sporulation.
abstract: Bacteria from the genus Streptomyces are among the most complex of all prokaryotes; not only do they grow as a complex mycelium, they also differentiate to form aerial hyphae before developing further to form spore chains. This developmental heterogeneity of streptomycete microcolonies makes studying the dynamic processes that contribute to growth and development a challenging procedure. As a result, in order to study the mechanisms that underpin streptomycete growth, we have developed a system for studying hyphal extension, protein trafficking, and sporulation by time-lapse microscopy. Through the use of time-lapse microscopy we have demonstrated that Streptomyces coelicolor germ tubes undergo a temporary arrest in their growth when in close proximity to sibling extension sites. Following germination, in this system, hyphae extended at a rate of approximately 20 microm h(-1), which was not significantly different from the rate at which the apical ring of the cytokinetic protein FtsZ progressed along extending hyphae through a spiraling movement. Although we were able to generate movies for streptomycete sporulation, we were unable to do so for either the erection of aerial hyphae or the early stages of sporulation. Despite this, it was possible to demonstrate an arrest of aerial hyphal development that we suggest is through the depolymerization of FtsZ-enhanced green fluorescent protein (GFP). Consequently, the imaging system reported here provides a system that allows the dynamic movement of GFP-tagged proteins involved in growth and development of S. coelicolor to be tracked and their role in cytokinesis to be characterized during the streptomycete life cycle.
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Science, University of Strathclyde, Royal College, Glasgow, United Kingdom.
authors.forename: Vinod
authors.initials: V
authors.lastname: Jyothikumar
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Tilley
authors.forename: Rashmi
authors.initials: R
authors.lastname: Wali
authors.forename: Paul R
authors.initials: PR
authors.lastname: Herron
mesh_terms: D019677:Artificial Gene Fusion; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D018715:Microscopy, Video / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000166:cytology* / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
references.citation: J Bacteriol. 1990 Jul;172(7):3637-43
references.pmid: 2113908
references.citation: J Gen Microbiol. 1978 Apr;105(2):203-14
references.pmid: 347027
references.citation: J Gen Microbiol. 1985 Apr;131(4):833-47
references.pmid: 3157776
references.citation: Mol Microbiol. 2006 Feb;59(3):731-42
references.pmid: 16420347
references.citation: Cell. 2003 Jun 13;113(6):767-76
references.pmid: 12809607
references.citation: Mol Microbiol. 2003 Feb;47(3):645-56
references.pmid: 12535067
references.citation: Environ Microbiol. 2007 Dec;9(12):3143-9
references.pmid: 17991041
references.citation: Mol Microbiol. 2007 Jun;64(5):1244-59
references.pmid: 17542918
references.citation: Trends Microbiol. 2006 Jul;14(7):313-9
references.pmid: 16759865
references.citation: Mol Microbiol. 2006 May;60(4):838-52
references.pmid: 16677297
references.citation: J Bacteriol. 1987 Nov;169(11):5002-7
references.pmid: 3117770
references.citation: Z Allg Mikrobiol. 1982;22(5):335-47
references.pmid: 7124001
references.citation: Mol Microbiol. 1994 Oct;14(2):243-54
references.pmid: 7830569
references.citation: Cell. 2008 Apr 4;133(1):90-102
references.pmid: 18394992
references.citation: J Bacteriol. 2001 Sep;183(17):5180-6
references.pmid: 11489872
references.citation: Curr Opin Microbiol. 2003 Dec;6(6):564-71
references.pmid: 14662351
references.citation: J Gen Microbiol. 1990 Jun;136(6):1077-84
references.pmid: 1696612
references.citation: Mol Microbiol. 1997 Sep;25(5):847-58
references.pmid: 9364911
references.citation: FEMS Microbiol Lett. 1992 Dec 15;100(1-3):351-9
references.pmid: 1478470
references.citation: Mol Microbiol. 1999 Dec;34(5):878-86
references.pmid: 10594815
references.citation: J Gen Microbiol. 1987 Aug;133(8):2089-96
references.pmid: 3443853
references.citation: Fungal Genet Biol. 2002 Oct;37(1):109-19
references.pmid: 12223195
references.citation: FEMS Microbiol Lett. 1999 Dec 1;181(1):159-63
references.pmid: 10564802
references.citation: J Bacteriol. 2005 May;187(9):3227-37
references.pmid: 15838050