Kalirin Interacts with TRAPP and Regulates Rab11 and Endosomal Recycling

• Published in: Cells (Basel, Switzerland) Vol. 9; no. 5; p. 1132
• Main Authors: Wang, Xiaolong, Weng, Meiqian, Ke, Yuting, Sapp, Ellen, DiFiglia, Marian, Li, Xueyi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 04.05.2020; MDPI
• Subjects: Animals; Cell adhesion & migration; Cell division; Cell membranes; Cold; Cytoskeleton; Deoxyribonucleic acid; DNA; Endocytosis; Endosomes; Endosomes - metabolism; Gene Knockdown Techniques; Guanine; Guanine nucleotide exchange factor; Guanine Nucleotide Exchange Factors - metabolism; HEK293 Cells; Humans; kalirin; Localization; Mass spectrometry; Mass spectroscopy; Membranes; Mice; Nerve Tissue Proteins - metabolism; Organelles; Plasmids; Protein Binding; Protein transport; Protein turnover; Proteins; rab GTP-Binding Proteins - metabolism; Rab11; Rac1 protein; Rats; Rho; Ribonucleic acid; RNA; Scientific imaging; TRAPP; Vesicular Transport Proteins - metabolism; kalirin; Rho; Rab11; guanine nucleotide exchange factor; TRAPP
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells9051132

Coordinated actions of Rab and Rho are necessary for numerous essential cellular processes ranging from vesicle budding to whole cell movement. How Rab and Rho are choreographed is poorly understood. Here, we report a protein complex comprised of kalirin, a Rho guanine nucleotide exchange factor (GEF) activating Rac1, and RabGEF transport protein particle (TRAPP). Kalirin was identified in a mass spectrometry analysis of proteins precipitated by trappc4 and detected on membranous organelles containing trappc4. Acute knockdown of kalirin did not affect trappc4, but significantly reduced overall and membrane-bound levels of trappc9, which specifies TRAPP toward activating Rab11. Trappc9 deficiency led to elevated expression of kalirin in neurons. Co-localization of kalirin and Rab11 occurred at a low frequency in NRK cells under steady state and was enhanced upon expressing an inactive Rab11 mutant to prohibit the dissociation of Rab11 from the kalirin-TRAPP complex. The small RNA-mediated depletion of kalirin diminished activities in cellular membranes for activating Rab11 and resulted in a shift in size of Rab11 positive structures from small to larger ones and tubulation of recycling endosomes. Our study suggests that kalirin and TRAPP form a dual GEF complex to choreograph actions of Rab11 and Rac1 at recycling endosomes.