Casein kinase II phosphorylation of cyclin F at serine 621 regulates the Lys48-ubiquitylation E3 ligase activity of the SCF(cyclin F) complex

Albert Lee, Stephanie L. Rayner, Alana De Luca, Serene S. L. Gwee, Marco Morsch, Vinod Sundaramoorthy, Hamideh Shahheydari, Audrey Ragagnin, Bingyang Shi, Shu Yang, Kelly L. Williams, Emily K. Don, Adam K. Walker, Katharine Y. Zhang, Justin J. Yerbury, Nicholas J. Cole, Julie D. Atkin, Ian P. Blair, Mark P. Molloy, Roger S. Chung

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder that is characterized by progressive weakness, paralysis and muscle loss often resulting in patient death within 3 – 5 years of diagnosis. Recently, we identified disease-linked mutations in the CCNF gene, which encodes the cyclin F protein, in cohorts of patients with familial and sporadic ALS and frontotemporal dementia (FTD) (Williams KL et al. 2016 Nat. Commun. 7, 11253. (doi:10.1038/ ncomms11253)). Cyclin F is a part of a Skp1-Cul-F-box (SCF) E3 ubiquitin-protein ligase complex and is responsible for ubiquitylating proteins for degradation by the proteasome. In this study, we investigated the phosphorylation status of cyclin F and the effect of the serine to glycine substitution at site 621 (S621G) on E3 ligase activity. This specific mutation (S621G) was found in a multi-generational Australian family with ALS/FTD. We identified seven phosphorylation sites on cyclin F, of which five are newly reported including Ser621. These phosphorylation sites were mostly identified within the PEST (proline, glutamic acid, serine and threonine) sequence located at the C-terminus of cyclin F. Additionally, we determined that casein kinase II (CK2) can phosphorylate Ser621 and thereby regulate the E3 ligase activity of the SCF(cyclin F) complex. Furthermore, the S621G mutation in cyclin F prevents phosphorylation by CK2 and confers elevated Lys48-ubiquitylation activity, a hallmark of ALS/FTD pathology. These findings highlight the importance of phosphorylation in regulating the activity of the SCF(cyclin F) E3 ligase complex that can affect downstream processes and may lead to defective motor neuron development, neuron degeneration and ultimately ALS and FTD.

LanguageEnglish
Article number170058
Pages1-11
Number of pages11
JournalOpen Biology
Volume7
Issue number10
DOIs
Publication statusPublished - 2017

Fingerprint

Casein Kinase II
Phosphorylation
Cyclins
Ubiquitin-Protein Ligases
Ubiquitination
Serine
Amyotrophic Lateral Sclerosis
Ubiquitin-Protein Ligase Complexes
Mutation
Cyclin C
Neurons
Frontotemporal Dementia
Nerve Degeneration
Muscle Weakness
Motor Neurons
Proteasome Endopeptidase Complex
Threonine
Proline
Paralysis
Neurodegenerative Diseases

Bibliographical note

Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • Amyotrophic lateral sclerosis
  • CCNF
  • Cyclin F
  • Frontotemporal dementia
  • Phosphorylation
  • Ubiquitylation

Cite this

@article{9ce9a485d0c640e79ee3933a918e5b62,
title = "Casein kinase II phosphorylation of cyclin F at serine 621 regulates the Lys48-ubiquitylation E3 ligase activity of the SCF(cyclin F) complex",
abstract = "Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder that is characterized by progressive weakness, paralysis and muscle loss often resulting in patient death within 3 – 5 years of diagnosis. Recently, we identified disease-linked mutations in the CCNF gene, which encodes the cyclin F protein, in cohorts of patients with familial and sporadic ALS and frontotemporal dementia (FTD) (Williams KL et al. 2016 Nat. Commun. 7, 11253. (doi:10.1038/ ncomms11253)). Cyclin F is a part of a Skp1-Cul-F-box (SCF) E3 ubiquitin-protein ligase complex and is responsible for ubiquitylating proteins for degradation by the proteasome. In this study, we investigated the phosphorylation status of cyclin F and the effect of the serine to glycine substitution at site 621 (S621G) on E3 ligase activity. This specific mutation (S621G) was found in a multi-generational Australian family with ALS/FTD. We identified seven phosphorylation sites on cyclin F, of which five are newly reported including Ser621. These phosphorylation sites were mostly identified within the PEST (proline, glutamic acid, serine and threonine) sequence located at the C-terminus of cyclin F. Additionally, we determined that casein kinase II (CK2) can phosphorylate Ser621 and thereby regulate the E3 ligase activity of the SCF(cyclin F) complex. Furthermore, the S621G mutation in cyclin F prevents phosphorylation by CK2 and confers elevated Lys48-ubiquitylation activity, a hallmark of ALS/FTD pathology. These findings highlight the importance of phosphorylation in regulating the activity of the SCF(cyclin F) E3 ligase complex that can affect downstream processes and may lead to defective motor neuron development, neuron degeneration and ultimately ALS and FTD.",
keywords = "Amyotrophic lateral sclerosis, CCNF, Cyclin F, Frontotemporal dementia, Phosphorylation, Ubiquitylation",
author = "Albert Lee and Rayner, {Stephanie L.} and {De Luca}, Alana and Gwee, {Serene S. L.} and Marco Morsch and Vinod Sundaramoorthy and Hamideh Shahheydari and Audrey Ragagnin and Bingyang Shi and Shu Yang and Williams, {Kelly L.} and Don, {Emily K.} and Walker, {Adam K.} and Zhang, {Katharine Y.} and Yerbury, {Justin J.} and Cole, {Nicholas J.} and Atkin, {Julie D.} and Blair, {Ian P.} and Molloy, {Mark P.} and Chung, {Roger S.}",
note = "Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.",
year = "2017",
doi = "10.1098/rsob.170058",
language = "English",
volume = "7",
pages = "1--11",
journal = "Open Biology",
issn = "2046-2441",
publisher = "Royal Society Publishing",
number = "10",

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TY - JOUR

T1 - Casein kinase II phosphorylation of cyclin F at serine 621 regulates the Lys48-ubiquitylation E3 ligase activity of the SCF(cyclin F) complex

AU - Lee, Albert

AU - Rayner, Stephanie L.

AU - De Luca, Alana

AU - Gwee, Serene S. L.

AU - Morsch, Marco

AU - Sundaramoorthy, Vinod

AU - Shahheydari, Hamideh

AU - Ragagnin, Audrey

AU - Shi, Bingyang

AU - Yang, Shu

AU - Williams, Kelly L.

AU - Don, Emily K.

AU - Walker, Adam K.

AU - Zhang, Katharine Y.

AU - Yerbury, Justin J.

AU - Cole, Nicholas J.

AU - Atkin, Julie D.

AU - Blair, Ian P.

AU - Molloy, Mark P.

AU - Chung, Roger S.

N1 - Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

PY - 2017

Y1 - 2017

N2 - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder that is characterized by progressive weakness, paralysis and muscle loss often resulting in patient death within 3 – 5 years of diagnosis. Recently, we identified disease-linked mutations in the CCNF gene, which encodes the cyclin F protein, in cohorts of patients with familial and sporadic ALS and frontotemporal dementia (FTD) (Williams KL et al. 2016 Nat. Commun. 7, 11253. (doi:10.1038/ ncomms11253)). Cyclin F is a part of a Skp1-Cul-F-box (SCF) E3 ubiquitin-protein ligase complex and is responsible for ubiquitylating proteins for degradation by the proteasome. In this study, we investigated the phosphorylation status of cyclin F and the effect of the serine to glycine substitution at site 621 (S621G) on E3 ligase activity. This specific mutation (S621G) was found in a multi-generational Australian family with ALS/FTD. We identified seven phosphorylation sites on cyclin F, of which five are newly reported including Ser621. These phosphorylation sites were mostly identified within the PEST (proline, glutamic acid, serine and threonine) sequence located at the C-terminus of cyclin F. Additionally, we determined that casein kinase II (CK2) can phosphorylate Ser621 and thereby regulate the E3 ligase activity of the SCF(cyclin F) complex. Furthermore, the S621G mutation in cyclin F prevents phosphorylation by CK2 and confers elevated Lys48-ubiquitylation activity, a hallmark of ALS/FTD pathology. These findings highlight the importance of phosphorylation in regulating the activity of the SCF(cyclin F) E3 ligase complex that can affect downstream processes and may lead to defective motor neuron development, neuron degeneration and ultimately ALS and FTD.

AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder that is characterized by progressive weakness, paralysis and muscle loss often resulting in patient death within 3 – 5 years of diagnosis. Recently, we identified disease-linked mutations in the CCNF gene, which encodes the cyclin F protein, in cohorts of patients with familial and sporadic ALS and frontotemporal dementia (FTD) (Williams KL et al. 2016 Nat. Commun. 7, 11253. (doi:10.1038/ ncomms11253)). Cyclin F is a part of a Skp1-Cul-F-box (SCF) E3 ubiquitin-protein ligase complex and is responsible for ubiquitylating proteins for degradation by the proteasome. In this study, we investigated the phosphorylation status of cyclin F and the effect of the serine to glycine substitution at site 621 (S621G) on E3 ligase activity. This specific mutation (S621G) was found in a multi-generational Australian family with ALS/FTD. We identified seven phosphorylation sites on cyclin F, of which five are newly reported including Ser621. These phosphorylation sites were mostly identified within the PEST (proline, glutamic acid, serine and threonine) sequence located at the C-terminus of cyclin F. Additionally, we determined that casein kinase II (CK2) can phosphorylate Ser621 and thereby regulate the E3 ligase activity of the SCF(cyclin F) complex. Furthermore, the S621G mutation in cyclin F prevents phosphorylation by CK2 and confers elevated Lys48-ubiquitylation activity, a hallmark of ALS/FTD pathology. These findings highlight the importance of phosphorylation in regulating the activity of the SCF(cyclin F) E3 ligase complex that can affect downstream processes and may lead to defective motor neuron development, neuron degeneration and ultimately ALS and FTD.

KW - Amyotrophic lateral sclerosis

KW - CCNF

KW - Cyclin F

KW - Frontotemporal dementia

KW - Phosphorylation

KW - Ubiquitylation

UR - http://www.scopus.com/inward/record.url?scp=85037045093&partnerID=8YFLogxK

U2 - 10.1098/rsob.170058

DO - 10.1098/rsob.170058

M3 - Article

VL - 7

SP - 1

EP - 11

JO - Open Biology

T2 - Open Biology

JF - Open Biology

SN - 2046-2441

IS - 10

M1 - 170058

ER -