Localization of protein kinase NDR2 to peroxisomes and its role in ciliogenesis

Shoko Abe, Tomoaki Nagai, Moe Masukawa, Kanji Okumoto, Yuta Homma, Yukio Fujiki, Kensaku Mizuno

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Nuclear Dbf2-related (NDR) kinases, comprising NDR1 and NDR2, are serine/threonine kinases that play crucial roles in the control of cell proliferation, apoptosis, and morphogenesis. We recently showed that NDR2, but not NDR1, is involved in primary cilium formation; however, the mechanism underlying their functional difference in ciliogenesis is unknown. To address this issue, we examined their subcellular localization. Despite their close sequence similarity, NDR2 exhibited punctate localization in the cytoplasm, whereas NDR1 was diffusely distributed within the cell. Notably, NDR2 puncta mostly co-localized with the peroxisome marker proteins, catalase and CFP-SKL (cyan fluorescent protein carrying the C-terminal typical peroxisome-targeting signal type-1 (PTS1) sequence, Ser-Lys-Leu). NDR2 contains the PTS1-like sequence, Gly-Lys-Leu, at the C-terminal end, whereas the C-terminal end of NDR1 is Ala-Lys. An NDR2 mutant lacking the C-terminal Leu, NDR2(ΔL), exhibited almost diffuse distribution in cells. Additionally, NDR2, but neither NDR1 nor NDR2(ΔL), bound to the PTS1 receptor Pex5p. Together, these findings indicate that NDR2 localizes to the peroxisome by using the C-terminal GKL sequence. Intriguingly, topology analysis of NDR2 suggests that NDR2 is exposed to the cytosolic surface of the peroxisome. The expression of wild-type NDR2, but not NDR2(ΔL), recovered the suppressive effect of NDR2 knockdown on ciliogenesis. Furthermore, knockdown of peroxisome biogenesis factor genes (PEX1 or PEX3) partially suppressed ciliogenesis. These results suggest that the peroxisomal localization of NDR2 is implicated in its function to promote primary cilium formation.

Original languageEnglish
Pages (from-to)4089-4098
Number of pages10
JournalJournal of Biological Chemistry
Volume292
Issue number10
DOIs
Publication statusPublished - Mar 10 2017

Fingerprint

Peroxisomes
Protein-Serine-Threonine Kinases
Cell proliferation
Catalase
Protein Kinases
Phosphotransferases
Genes
Topology
Apoptosis
Proteins
Cilia
seryl-lysyl-leucine
Protein Sorting Signals
Protein C
Morphogenesis
Cyan Fluorescent Protein
peroxisome-targeting signal 1 receptor
Cytoplasm
Cell Proliferation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Localization of protein kinase NDR2 to peroxisomes and its role in ciliogenesis. / Abe, Shoko; Nagai, Tomoaki; Masukawa, Moe; Okumoto, Kanji; Homma, Yuta; Fujiki, Yukio; Mizuno, Kensaku.

In: Journal of Biological Chemistry, Vol. 292, No. 10, 10.03.2017, p. 4089-4098.

Research output: Contribution to journalArticle

Abe, S, Nagai, T, Masukawa, M, Okumoto, K, Homma, Y, Fujiki, Y & Mizuno, K 2017, 'Localization of protein kinase NDR2 to peroxisomes and its role in ciliogenesis', Journal of Biological Chemistry, vol. 292, no. 10, pp. 4089-4098. https://doi.org/10.1074/jbc.M117.775916
Abe, Shoko ; Nagai, Tomoaki ; Masukawa, Moe ; Okumoto, Kanji ; Homma, Yuta ; Fujiki, Yukio ; Mizuno, Kensaku. / Localization of protein kinase NDR2 to peroxisomes and its role in ciliogenesis. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 10. pp. 4089-4098.
@article{31d2382d6cec4dd19533acb4b04c5690,
title = "Localization of protein kinase NDR2 to peroxisomes and its role in ciliogenesis",
abstract = "Nuclear Dbf2-related (NDR) kinases, comprising NDR1 and NDR2, are serine/threonine kinases that play crucial roles in the control of cell proliferation, apoptosis, and morphogenesis. We recently showed that NDR2, but not NDR1, is involved in primary cilium formation; however, the mechanism underlying their functional difference in ciliogenesis is unknown. To address this issue, we examined their subcellular localization. Despite their close sequence similarity, NDR2 exhibited punctate localization in the cytoplasm, whereas NDR1 was diffusely distributed within the cell. Notably, NDR2 puncta mostly co-localized with the peroxisome marker proteins, catalase and CFP-SKL (cyan fluorescent protein carrying the C-terminal typical peroxisome-targeting signal type-1 (PTS1) sequence, Ser-Lys-Leu). NDR2 contains the PTS1-like sequence, Gly-Lys-Leu, at the C-terminal end, whereas the C-terminal end of NDR1 is Ala-Lys. An NDR2 mutant lacking the C-terminal Leu, NDR2(ΔL), exhibited almost diffuse distribution in cells. Additionally, NDR2, but neither NDR1 nor NDR2(ΔL), bound to the PTS1 receptor Pex5p. Together, these findings indicate that NDR2 localizes to the peroxisome by using the C-terminal GKL sequence. Intriguingly, topology analysis of NDR2 suggests that NDR2 is exposed to the cytosolic surface of the peroxisome. The expression of wild-type NDR2, but not NDR2(ΔL), recovered the suppressive effect of NDR2 knockdown on ciliogenesis. Furthermore, knockdown of peroxisome biogenesis factor genes (PEX1 or PEX3) partially suppressed ciliogenesis. These results suggest that the peroxisomal localization of NDR2 is implicated in its function to promote primary cilium formation.",
author = "Shoko Abe and Tomoaki Nagai and Moe Masukawa and Kanji Okumoto and Yuta Homma and Yukio Fujiki and Kensaku Mizuno",
year = "2017",
month = "3",
day = "10",
doi = "10.1074/jbc.M117.775916",
language = "English",
volume = "292",
pages = "4089--4098",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "10",

}

TY - JOUR

T1 - Localization of protein kinase NDR2 to peroxisomes and its role in ciliogenesis

AU - Abe, Shoko

AU - Nagai, Tomoaki

AU - Masukawa, Moe

AU - Okumoto, Kanji

AU - Homma, Yuta

AU - Fujiki, Yukio

AU - Mizuno, Kensaku

PY - 2017/3/10

Y1 - 2017/3/10

N2 - Nuclear Dbf2-related (NDR) kinases, comprising NDR1 and NDR2, are serine/threonine kinases that play crucial roles in the control of cell proliferation, apoptosis, and morphogenesis. We recently showed that NDR2, but not NDR1, is involved in primary cilium formation; however, the mechanism underlying their functional difference in ciliogenesis is unknown. To address this issue, we examined their subcellular localization. Despite their close sequence similarity, NDR2 exhibited punctate localization in the cytoplasm, whereas NDR1 was diffusely distributed within the cell. Notably, NDR2 puncta mostly co-localized with the peroxisome marker proteins, catalase and CFP-SKL (cyan fluorescent protein carrying the C-terminal typical peroxisome-targeting signal type-1 (PTS1) sequence, Ser-Lys-Leu). NDR2 contains the PTS1-like sequence, Gly-Lys-Leu, at the C-terminal end, whereas the C-terminal end of NDR1 is Ala-Lys. An NDR2 mutant lacking the C-terminal Leu, NDR2(ΔL), exhibited almost diffuse distribution in cells. Additionally, NDR2, but neither NDR1 nor NDR2(ΔL), bound to the PTS1 receptor Pex5p. Together, these findings indicate that NDR2 localizes to the peroxisome by using the C-terminal GKL sequence. Intriguingly, topology analysis of NDR2 suggests that NDR2 is exposed to the cytosolic surface of the peroxisome. The expression of wild-type NDR2, but not NDR2(ΔL), recovered the suppressive effect of NDR2 knockdown on ciliogenesis. Furthermore, knockdown of peroxisome biogenesis factor genes (PEX1 or PEX3) partially suppressed ciliogenesis. These results suggest that the peroxisomal localization of NDR2 is implicated in its function to promote primary cilium formation.

AB - Nuclear Dbf2-related (NDR) kinases, comprising NDR1 and NDR2, are serine/threonine kinases that play crucial roles in the control of cell proliferation, apoptosis, and morphogenesis. We recently showed that NDR2, but not NDR1, is involved in primary cilium formation; however, the mechanism underlying their functional difference in ciliogenesis is unknown. To address this issue, we examined their subcellular localization. Despite their close sequence similarity, NDR2 exhibited punctate localization in the cytoplasm, whereas NDR1 was diffusely distributed within the cell. Notably, NDR2 puncta mostly co-localized with the peroxisome marker proteins, catalase and CFP-SKL (cyan fluorescent protein carrying the C-terminal typical peroxisome-targeting signal type-1 (PTS1) sequence, Ser-Lys-Leu). NDR2 contains the PTS1-like sequence, Gly-Lys-Leu, at the C-terminal end, whereas the C-terminal end of NDR1 is Ala-Lys. An NDR2 mutant lacking the C-terminal Leu, NDR2(ΔL), exhibited almost diffuse distribution in cells. Additionally, NDR2, but neither NDR1 nor NDR2(ΔL), bound to the PTS1 receptor Pex5p. Together, these findings indicate that NDR2 localizes to the peroxisome by using the C-terminal GKL sequence. Intriguingly, topology analysis of NDR2 suggests that NDR2 is exposed to the cytosolic surface of the peroxisome. The expression of wild-type NDR2, but not NDR2(ΔL), recovered the suppressive effect of NDR2 knockdown on ciliogenesis. Furthermore, knockdown of peroxisome biogenesis factor genes (PEX1 or PEX3) partially suppressed ciliogenesis. These results suggest that the peroxisomal localization of NDR2 is implicated in its function to promote primary cilium formation.

UR - http://www.scopus.com/inward/record.url?scp=85015073601&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85015073601&partnerID=8YFLogxK

U2 - 10.1074/jbc.M117.775916

DO - 10.1074/jbc.M117.775916

M3 - Article

C2 - 28122914

AN - SCOPUS:85015073601

VL - 292

SP - 4089

EP - 4098

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 10

ER -