Harden N, Lee J, Loh H Y, Ong Y M, Tan I, Leung T, Manser E, Lim L
Glaxo-IMCB Group, Institute of Molecular and Cell Biology, National University of Singapore.
Mol Cell Biol. 1996 May;16(5):1896-908. doi: 10.1128/MCB.16.5.1896.
Changes in cell morphology are essential in the development of a multicellular organism. The regulation of the cytoskeleton by the Rho subfamily of small GTP-binding proteins is an important determinant of cell shape. The Rho subfamily has been shown to participate in a variety of morphogenetic processes during Drosophila melanogaster development. We describe here a Drosophila homolog, DPAK, of the serine/threonine kinase PAK, a protein which is a target of the Rho subfamily proteins Rac and Cdc42. Rac, Cdc42, and PAK have previously been implicated in signaling by c-Jun amino-terminal kinases. DPAK bound to activated (GTP-bound) Drosophila Rac (DRacA) and Drosophila Cdc42. Similarities in the distributions of DPAK, integrin, and phosphotyrosine suggested an association of DPAK with focal adhesions and Cdc42- and Rac-induced focal adhesion-like focal complexes. DPAK was elevated in the leading edge of epidermal cells, whose morphological changes drive dorsal closure of the embryo. We have previously shown that the accumulation of cytoskeletal elements initiating cell shape changes in these cells could be inhibited by expression of a dominant-negative DRacA transgene. We show that leading-edge epidermal cells flanking segment borders, which express particularly large amounts of DPAK, undergo transient losses of cytoskeletal structures during dorsal closure. We propose that DPAK may be regulating the cytoskeleton through its association with focal adhesions and focal complexes and may be participating with DRacA in a c-Jun amino-terminal kinase signaling pathway recently demonstrated to be required for dorsal closure.
细胞形态的变化在多细胞生物体的发育过程中至关重要。小GTP结合蛋白Rho亚家族对细胞骨架的调控是细胞形状的一个重要决定因素。已表明Rho亚家族在黑腹果蝇发育过程中参与多种形态发生过程。我们在此描述了丝氨酸/苏氨酸激酶PAK的果蝇同源物DPAK,PAK是Rho亚家族蛋白Rac和Cdc42的作用靶点。Rac、Cdc42和PAK先前已被证明参与c-Jun氨基末端激酶的信号传导。DPAK与活化的(GTP结合的)果蝇Rac(DRacA)和果蝇Cdc42结合。DPAK、整合素和磷酸酪氨酸分布的相似性表明DPAK与粘着斑以及Cdc42和Rac诱导的粘着斑样粘着复合体有关。DPAK在表皮细胞的前缘升高,其形态变化驱动胚胎的背侧闭合。我们先前已表明,在这些细胞中引发细胞形状变化的细胞骨架成分的积累可被显性负性DRacA转基因的表达所抑制。我们发现,在背侧闭合过程中,位于节段边界两侧且大量表达DPAK的前缘表皮细胞会经历细胞骨架结构的短暂丧失。我们提出,DPAK可能通过与粘着斑和粘着复合体的结合来调节细胞骨架,并且可能与DRacA一起参与最近证明对背侧闭合必需的c-Jun氨基末端激酶信号通路。