TY - JOUR
T1 - Quantitative insights into actin rearrangements and bacterial target site selection from SalmonellaTyphimurium infection of micropatterned cells
AU - Vonaesch, Pascale
AU - Cardini, Steven
AU - Sellin, Mikael E.
AU - Goud, Bruno
AU - Hardt, Wolf Dietrich
AU - Schauer, Kristine
PY - 2013/11/1
Y1 - 2013/11/1
N2 - Summary: Reorganization of the host cell actin cytoskeleton is crucial during pathogen invasion. We established micropatterned cells as a standardized infection model for cell invasion to quantitatively study actin rearrangements triggered by SalmonellaTyphimurium (S. Tm). Micropatterns of extracellular matrix proteins force cells to adopt a reproducible shape avoiding strong cell-to-cell variations, a major limitation in classical cell culture conditions. S.Tm induced F-actin-rich ruffles and invaded micropatterned cells similar to unconstrained cells. Yet, standardized conditions allowed fast and unbiased comparison of cellular changes triggered by the SipA and SopE bacterial effector proteins. Intensity measurements in defined regions revealed that the content of pre-existing F-actin remained unchanged during infection, suggesting that newly polymerized F-actin in bacteria-triggered ruffles originates from the G-actin pool. Analysing bacterial target sites, we found that bacteria did not show any preferences for the local actin cytoskeleton specificities. Rather, invasion was constrained to a specific 'cell height', due to flagella-mediated near-surface swimming. We found that invasion sites were similar to bacterial binding sites, indicating that S. Tm can induce a permissive invasion site wherever it binds. As micropatterned cells can be infected by many different pathogens they represent a valuable new tool for quantitative analysis of host-pathogen interactions.
AB - Summary: Reorganization of the host cell actin cytoskeleton is crucial during pathogen invasion. We established micropatterned cells as a standardized infection model for cell invasion to quantitatively study actin rearrangements triggered by SalmonellaTyphimurium (S. Tm). Micropatterns of extracellular matrix proteins force cells to adopt a reproducible shape avoiding strong cell-to-cell variations, a major limitation in classical cell culture conditions. S.Tm induced F-actin-rich ruffles and invaded micropatterned cells similar to unconstrained cells. Yet, standardized conditions allowed fast and unbiased comparison of cellular changes triggered by the SipA and SopE bacterial effector proteins. Intensity measurements in defined regions revealed that the content of pre-existing F-actin remained unchanged during infection, suggesting that newly polymerized F-actin in bacteria-triggered ruffles originates from the G-actin pool. Analysing bacterial target sites, we found that bacteria did not show any preferences for the local actin cytoskeleton specificities. Rather, invasion was constrained to a specific 'cell height', due to flagella-mediated near-surface swimming. We found that invasion sites were similar to bacterial binding sites, indicating that S. Tm can induce a permissive invasion site wherever it binds. As micropatterned cells can be infected by many different pathogens they represent a valuable new tool for quantitative analysis of host-pathogen interactions.
UR - http://www.scopus.com/inward/record.url?scp=84885433863&partnerID=8YFLogxK
U2 - 10.1111/cmi.12154
DO - 10.1111/cmi.12154
M3 - Article
C2 - 23648178
AN - SCOPUS:84885433863
SN - 1462-5814
VL - 15
SP - 1851
EP - 1865
JO - Cellular Microbiology
JF - Cellular Microbiology
IS - 11
ER -