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PMID:12952073

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Citation

Müller-Taubenberger, A, Bretschneider, T, Faix, J, Konzok, A, Simmeth, E and Weber, I (2002) Differential localization of the Dictyostelium kinase DPAKa during cytokinesis and cell migration. J. Muscle Res. Cell. Motil. 23:751-63

Abstract

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.

Links

PubMed

Keywords

Animals; Cell Division/physiology; Chemotaxis/physiology; Dictyostelium/cytology; Dictyostelium/enzymology; Dictyostelium/physiology; Genes, Reporter; Green Fluorescent Proteins; Luminescent Proteins/genetics; Luminescent Proteins/metabolism; Movement/physiology; Protein-Serine-Threonine Kinases/genetics; Protein-Serine-Threonine Kinases/metabolism; Recombinant Fusion Proteins/metabolism; p21-Activated Kinases

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DICDI:RAC1A

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q55D99

F

Seeded From UniProt

complete

DICDI:PAKA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P34144

F

Seeded From UniProt

complete

DICDI:PAKA

located_in

GO:0031254: cell trailing edge

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:PAKA

located_in

GO:0097204: phagocytic cup base

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:PAKA

NOT|located_in

GO:0032154: cleavage furrow

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:PAKA

acts_upstream_of_or_within

GO:0006909: phagocytosis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:PAKA

located_in

GO:0005938: cell cortex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:PAKA

acts_upstream_of_or_within

GO:0048870: cell motility

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:MYS2

part_of

GO:0097204: phagocytic cup base

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

Notes

See also

References

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