Differential localization of the Dictyostelium kinase DPAKa during cytokinesis and cell migration.
The Dictyostelium kinase DPAKa is a member of the p21-activated kinase (PAK) family, consisting of an N-terminal domain characterized by a coiled-coil region and proline-rich motifs, a Rac-binding CRIB-domain, and a highly conserved C-terminal kinase domain. In this study we show that cells overexpressing a C-terminal DPAKa fragment comprising the kinase domain are significantly impaired in motility and phagocytosis, whereas DPAKa-null cells display no obvious phenotypic change. We analyzed the in vivo localization of full-length and truncated DPAKa tagged with green fluorescent protein (GFP). The N-terminal fragments show a highly dynamic cortical localization without a permanent polarized enrichment, whereas the C-terminal fragment is homogenously distributed throughout the cell. The localization of full-length DPAKa is similar to that of myosin II at the rear end of locomoting cells and at the base of phagocytic cups. During mitosis DPAKa is gradually recruited to the cell cortex starting at metaphase, which also parallels the dynamics of myosin II cortical recruitment. However, in contrast to myosin II, DPAKa does not accumulate in the cleavage furrow but stays uniformly distributed throughout the cell cortex. This finding contrasts with previous work claiming accumulation of DPAKa in the cleavage furrow of dividing cells. Our results suggest that the N-terminus directs DPAKa to the cortex, and the C-terminus is necessary for restricting its localization to the rear of moving cells during chemotaxis. Therefore, DPAKa may play distinct roles in myosin II regulation during cell movement and cell division.
Müller-Taubenberger
Annette
A
Max-Planck-Institut für Biochemie, 82152 Martinsried, Germany. amueller@biochem.mpg.de
Bretschneider
Till
T
Faix
Jan
J
Konzok
Angelika
A
Simmeth
Evelyn
E
Weber
Igor
I
D000818:Animals; D002455:Cell Division / Q000502:physiology*; D002633:Chemotaxis / Q000502:physiology; D004023:Dictyostelium / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009068:Movement / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D054462:p21-Activated Kinases
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases