Stathmin recruits tubulin to Listeria monocytogenes-induced actin comets and promotes bacterial dissemination
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The tubulin cytoskeleton is one of the main components of the cytoarchitecture and is involved in several cellular functions. Here, we examine the interplay between Listeria monocytogenes (Lm) and the tubulin cytoskeleton upon cellular infection. We show that non-polymeric tubulin is present throughout Lm actin comet tails and, to a less extent, in actin clouds. Moreover, we demonstrate that stathmin, a regulator of microtubule dynamics, is also found in these Lm-associated actin structures and is required for tubulin recruitment. Depletion of host stathmin results in longer comets containing less F-actin, which may be correlated with higher levels of inactive cofilin in the comet, thus suggesting a defect on local F-actin dynamics. In addition, intracellular bacterial speed is significantly reduced in stathmin-depleted cells, revealing the importance of stathmin/tubulin in intracellular Lm motility. In agreement, the area of infection foci and the total bacterial loads are also significantly reduced in stathmin-depleted cells. Collectively, our results demonstrate that stathmin promotes efficient cellular infection, possibly through tubulin recruitment and control of actin dynamics at Lm-polymerized actin structures.
KeywordsListeria monocytogenes Tubulin Stathmin Cytoskeleton Actin comet tails Infection
Multiplicity of infection
Stimulated emission depletion
Region of interest
Mean fluorescence intensity
Green fluorescent protein
This work received funding from Norte-01-0145-FEDER-000012—Structured program on bioengineered therapies for infectious diseases and tissue regeneration, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). ACC and FC were supported by a Fundação para a Ciência e Tecnologia (FCT) Post-doctoral Fellowship (SFRH/BPD/88769/2012) and Ph.D. fellowship (SFRH/BD/61825/2009), respectively, through FCT/MEC co-funded by QREN and POPH (Programa Operacional Potencial Humano). SS was supported by FCT Investigator program (COMPETE, POPH, and FCT). We thank ALM unit from IBMC/i3S for technical support, B. Fonseca (FFUP) for the BeWo cell line and H. Maiato, C. Sunkel and J. B. Relvas laboratories (IBMC/i3S) for sharing reagents. We are also thankful to J. Ferreira and A. Pereira for fruitful discussions.
ACC, DC and SS conceived and designed the experiments; ACC and FC performed the experiments; ACC, FC, DC and SS analyzed the data; DC and SS obtained the funding; ACC and SS wrote the manuscript.
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