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Originally published as MBC in Press, 10.1091/mbc.E07-07-0640 on December 27, 2007

Vol. 19, Issue 3, 1241-1251, March 2008

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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

Submitted July 9, 2007; Revised November 20, 2007; Accepted December 14, 2007
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 small interfering RNA 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. Therefore, these results show that in J774 cells coronin 1 has a specific role in modulating phagosome–lysosome transport upon mycobacterial infection and that it is dispensable for most F-actin–mediated cytoskeletal rearrangements.

This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E07-07-0640) on December 27, 2007.

* These authors contributed equally to this work.

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






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