Functional analysis of slac2-a/melanophilin as a linker protein between Rab27A and myosin Va in melanosome transport.
Slac2-a/melanophilin regulates melanosome transport in mammalian skin melanocytes by linking melanosome-bound Rab27A and an actin-based motor protein, myosin Va. Slac2-a consists of an N-terminal Slp homology domain (SHD), which has been identified as a specific GTP-Rab27-binding domain, a myosin Va-binding domain (MBD) in the middle region, and an actin-binding domain (ABD) at the C-terminus. Mutations in the slac2-a/mlph gene cause the abnormal pigmentation (i.e., perinuclear melanosome aggregation in melanocytes) in human Griscelli syndrome type III and in leaden mice because of the inability to form the tripartite protein complex consisting of Rab27A, Slac2-a, and myosin Va. In this chapter we describe the methods, including in vivo melanosome distribution assay combined with dominant-negative approaches and RNA interference technology, that have been used to analyze the function of Slac2-a in melanosome transport in melanocytes.
Taruho S
TS
Kuroda
Takashi
T
Itoh
Mitsunori
M
Fukuda
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000502:physiology*; D002478:Cells, Cultured; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D020460:Melanosomes / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018995:Myosin Heavy Chains / Q000502:physiology*; D024701:Myosin Type V / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering; D020691:rab GTP-Binding Proteins / Q000502:physiology*; D000075745:rab27 GTP-Binding Proteins
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D017931:DNA Primers; C435724:Mlph protein, mouse; C513082:Myo5a protein, mouse; D034741:RNA, Small Interfering; D000075745:rab27 GTP-Binding Proteins; D024701:Myosin Type V; C512962:Rab27a protein, mouse; D018995:Myosin Heavy Chains; D020691:rab GTP-Binding Proteins