Abr and Bcr, Two Homologous Rac GTPase-Activating Proteins, Control Multiple Cellular Functions of Murine Macrophages

Abstract

Small GTPases of the Rho family are key regulators of phagocytic leukocyte function. Abr and Bcr are homologous, multidomain proteins. Their C-terminal domain has GTPase-activating protein (GAP) activity that, in vitro, is specific for Rac and Cdc42. To address the in vivo relevance of these entire proteins, of which little is known, the current study examined the effect of the genetic ablation of Abr and Bcr in murine macrophages. The concomitant loss of Abr and Bcr induced multiple alterations of macrophage cellular behavior known to be under the control of Rac. Macrophages lacking both Abr and Bcr exhibited an atypical, elongated morphology that was reproduced by the ectopic expression of GAP domain mutant Abr and Bcr in a macrophage cell line and of constitutively active Rac in primary macrophages. A robust increase in colony-stimulating factor 1 (CSF-1)-directed motility was observed in macrophages deficient for both proteins and, in response to CSF-1 stimulation, Abr and Bcr transiently translocated to the plasma membrane. Phagocytosis of opsonized particles was also increased in macrophages lacking both proteins and correlated with sustained Rac activation. Bcr and Abr GAP mutant proteins localized around phagosomes and induced distinct phagocytic cup formation. These results identify Abr and Bcr as the only GAPs to date that specifically negatively regulate Rac function in vivo in primary macrophages.

SUPPLEMENTAL MATERIAL

We thank George McNamara for help with microscopy, Ana Romero for help with isolation of mouse tail DNAs, and Donna Foster for care of the mice. Gary Bokoch and Ulla Knaus (The Scripps Research Institute, La Jolla, CA) are acknowledged for GST-Pak-RBD, and JaeRan Lee and EunJoon Kim (Korea Advanced Institute of Science and Technology, Daejon, Korea) are acknowledged for the generous gift of the EGFP-Bcr R1090A- and EGFP-Abr R683A-encoding plasmids. We thank Allan Hall (Memorial Sloan-Kettering Cancer Center, New York) for Myc-V12Cdc42, Manabu Negishi (University of Kyoto, Kyoto, Japan) for Myc-RhoG and Myc-RhoG G12V, and Kodi Ravichandran (University of Virginia) for GST-ELMO1.

This work was supported by National Institutes of Health Public Health Service grants HL071945 and HL060231.