PLK1-dependent activation of LRRK1 regulates spindle orientation by phosphorylating CDK5RAP2

Hiroshi Hanafusa; Shin Kedashiro; Motohiro Tezuka; Motoki Funatsu; Satoshi Usami; Fumiko Toyoshima; Kunihiro Matsumoto

doi:10.1038/ncb3204

PLK1-dependent activation of LRRK1 regulates spindle orientation by phosphorylating CDK5RAP2

Nat Cell Biol. 2015 Aug;17(8):1024-35. doi: 10.1038/ncb3204. Epub 2015 Jul 20.

Authors

Hiroshi Hanafusa¹, Shin Kedashiro¹, Motohiro Tezuka¹, Motoki Funatsu¹, Satoshi Usami¹, Fumiko Toyoshima², Kunihiro Matsumoto¹

Affiliations

¹ Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan.
² Department of Cell Biology, Institute for Virus Research, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan.

PMID: 26192437
DOI: 10.1038/ncb3204

Abstract

Correct formation of the cell division axis requires the initial precise orientation of the mitotic spindle. Proper spindle orientation depends on centrosome maturation, and Polo-like kinase 1 (PLK1) is known to play a crucial role in this process. However, the molecular mechanisms that function downstream of PLK1 are not well understood. Here we show that LRRK1 is a PLK1 substrate that is phosphorylated on Ser 1790. PLK1 phosphorylation is required for CDK1-mediated activation of LRRK1 at the centrosomes, and this in turn regulates mitotic spindle orientation by nucleating the growth of astral microtubules from the centrosomes. Interestingly, LRRK1 in turn phosphorylates CDK5RAP2(Cep215), a human homologue of Drosophila Centrosomin (Cnn), in its γ-tubulin-binding motif, thus promoting the interaction of CDK5RAP2 with γ-tubulin. LRRK1 phosphorylation of CDK5RAP2 Ser 140 is necessary for CDK5RAP2-dependent microtubule nucleation. Thus, our findings provide evidence that LRRK1 regulates mitotic spindle orientation downstream of PLK1 through CDK5RAP2-dependent centrosome maturation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs
Animals
Binding Sites
CDC2 Protein Kinase
COS Cells
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism*
Centrosome / enzymology
Chlorocebus aethiops
Cyclin-Dependent Kinases / metabolism
Enzyme Activation
HEK293 Cells
HeLa Cells
Humans
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism*
Microtubules / enzymology
Mitosis*
Mutation
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism*
Phosphorylation
Polo-Like Kinase 1
Protein Binding
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism*
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins / metabolism*
RNA Interference
Serine
Signal Transduction
Spindle Apparatus / enzymology*
Time Factors
Transfection
Tubulin / metabolism

Substances

CDK5RAP2 protein, human
Cell Cycle Proteins
Intracellular Signaling Peptides and Proteins
Nerve Tissue Proteins
Proto-Oncogene Proteins
Tubulin
Serine
LRRK1 protein, human
Protein Serine-Threonine Kinases
CDC2 Protein Kinase
CDK1 protein, human
Cyclin-Dependent Kinases