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MBC in Press, published online ahead of print December 27, 2007
Mol. Biol. Cell 10.1091/mbc.E07-07-0640

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Submitted on July 9, 2007
Revised on November 20, 2007
Accepted on December 14, 2007

RNA Interference in J774 Macrophages Reveals a Role for Coronin 1 in Mycobacterial Trafficking but Not in Actin-dependent Processes

Rajesh Jayachandran,* John Gatfield,* Jan Massner, Imke Albrecht, Bettina Zanolari, and Jean Pieters

Biozentrum, University of Basel, 4056 Basel, Switzerland

Monitoring Editor: Sandra Schmid

Macrophages are crucial for innate immunity, apoptosis and tissue remodeling, processes that rely on the capacity of macrophages to internalize and process cargo through phagocytosis. Coronin 1, a member of the WD repeat protein family of coronins specifically expressed in leukocytes, was originally identified as a molecule that is recruited to mycobacterial phagosomes and prevents the delivery of mycobacteria to lysosomes, allowing these to survive within phagosomes. However, a role for coronin 1 in mycobacterial pathogenesis has been disputed in favor for its role in mediating phagocytosis and cell motility. In this study, a role for coronin 1 in actin-mediated cellular processes was addressed using RNA interference in the murine macrophage cell line J774. It is shown that the absence of coronin 1 in J774 macrophages expressing siRNA constructs specific for coronin 1 does not affect phagocytosis, macropinocytosis, cell locomotion or regulation of NADPH oxidase activity. However, in coronin 1-negative J774 cells, internalized mycobacteria were rapidly transferred to lysosomes and killed. These results therefore show that in J774 cells coronin 1 has a specific role in modulating phagosome-lysosome transport upon mycobacterial infection and is dispensable for most F-actin-mediated cytoskeletal rearrangements. Keywords: Coronin, macrophages, RNAi, phagocytosis, actin

*These authors contributed equally to this work.

Address correspondence to: Jean Pieters (jean.pieters{at}unibas.ch)






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