getting organized – how bacterial cells move proteins and dna anna buch 25.01.2010 martin...
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Getting organized –how bacterial cells move
proteins and DNA
Anna Buch25.01.2010
Martin Thanbichler and Lucy Shapiro
Nature Reviews, 2008
Model systems for bacterial cell biology
Box 1
• E. coli: history, genetic tools, physiology
• B. subtilis: cell differentiation, large size
• C. crescentus: cell division, synchonizable
mobile
sessile
• Diffusion and capture
Assembly of stationary protein complexes
SpoIVB
SpoIIQ
Figure 1
Mother cell
Phagocytosis-likeuptake
Septal membrane
• Targeted membrane insertion
Assembly of stationary protein complexes
SpoIVB
SpoIIIAH
SpoIIQ
Figure 1
• Targeted membrane insertion
Assembly of stationary protein complexes
Steinhauer et al., Mol Microbiol. 1999 32:367-77.; Pollard & Cooper, Science 2009 326:1208-12
IcsA: outer membrane protein, N-term is exposed to host cytoplasm
IcsP: Protease that cleaves off IcsA
Shigella flexneri:facultative intracellular pathogen
Dynamic protein scaffolds and cell shape:Bacterial actin-like cytoskeleton
Figure 2
Bundles of two or more protofilaments.
MreB dynamics in C. crescentus
Figure 2
MreB cables
Spiral like during growth
Ring-like during cell division
Architecture of MreB cables
Figure 2; Carballido-Lopez & Errington, Dev Cell. 2003 4:19-28.
B. subtilis, FRAP of GFP-Mbl
Regulation of cell-wall biosynthesis
Carballido-Lopez et al., Dev Cell. 2006 11:399-409
MreB homologues: MreBH and MblLytE: peptidoglycan hydrolase
Peptidoglycan (PG)synthetic machinery
PG-hydrolase subunit
CW binding subdomain
B. subtilis
Role of MreC in bacterial morphogenesis
Divakaruni et al., PNAS 2005 102:18602-7
C. Crescentus
PBC (penicillin-binding protein):
involved in peptidoglykan synthesis
MreC
DAPI
Crescentin
Ausmees et al., Cell. 2003 115:705-13.
C. crescentus:
creS::Tn5 -> no crescentin
creS::Tn5 + creS ->crescentin on plasmid
In-vitro assay
His-CreS filaments,
EM negative stain
Plasmid segregation
• Actin superfamiliy member (type II partitioning system)
• Walker ATPase (type I partitioning system)
• Tubulin homologue
Plasmid segregation by Walker-type ATPases
• Walker A cytoskeletal ATPase (WACA)
Adapted from Lim et al., PNAS 2005 102:17658-63
Plasmids F and pB171 of E. coli
TubZ: B. thuringiensis serovar israelensis (pBtoxis)
Plasmid segregation by a tubulin homologue
Larsen et al., Genes Dev. 2007 21:1340-52
E.coli, expressing TubZ-GFP, FRAP, time in sec
Model proposes treadmilling
Divisome: Bacterial cell-division apparatus
Allard & Cytrynbaum PNAS 2009 106:145-50; Erickson, PNAS 2009 106:9238-43
Z-ring: FitsZ filaments
Rod-shaped bacterium (e.g. E. coli)
Division-site placement: The Min system
Figure 5
minCDE operon:
MinD: WACA family
MinCD-complex: inhibit FtsZ-ring formation
MinE: represses MinCD activity
“Fail-safe mechanism”: nucleoid occlusion
B. subtilis: Noc
E. coli: SlmA
Division-site placement: The MipZ system
MipZ: ATPase, inhibits FtsZ-polymerization
ParB: chromosome partitioning protein
parS: cluster of sites, 15 kb away from ori
Conclusions
• Tubulin filaments:– cell-division apparatus, plasmid segregation
• Actin cables:– DNA partitioning, cell-shape determination,
protein localization
• WACA ATPases:– DNA segregation, cell-division plane
Outlook
• Positioning of proteins at cell poles– TipN– Peptidoglycans– Cardiolipin -> ProP
• Biochemical assembly mechanisms– Actin homologues– Tubulin homologues– WACA ATPases