Identification of a Calcium/Calmodulin-dependent Protein Kinase That Phosphorylates the Neurospora Circadian Clock Protein FREQUENCY

Yuhong Yang, Ping Cheng, Gang Zhi, Yi Liu

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Phosphorylation of circadian clock proteins represents a major regulatory step that controls circadian clocks. In Neurospora, the circadian clock protein FREQUENCY (FRQ) is progressively phosphorylated over time, and its level decreases when it is hyperphosphorylated. In this study, we showed that most of the kinase activity phosphorylating FRQ in vitro was calcium/ calmodulin-dependent, and the endogenous FRQ in the Neurospora extracts was phosphorylated by a Ca/ CaM-dependent kinase-like activity. From Neurospora cell extracts, an ∼50-kDa Ca/CaM-dependent kinase (CAMK-1) that can specifically phosphorylate FRQ was purified. In vitro, this kinase accounts for near half of the FRQ kinase activity, and it can phosphorylate the FRQ region that contains the three known functionally important phosphorylation sites. To understand the function of camk-I in vivo, it was disrupted in Neurospora by gene replacement. After germination from ascospores, the camk-1 null strains grew slowly, indicating that CAMK-1 plays an important role in growth and development of Neurospora. This phenotype was transient however, revealing redundancy in the system. Analysis of the camk-1 null strain revealed that the deletion of camk-1 affected phase, period, and light-induced phase shifting of the circadian conidiation rhythm. Taken together, our results suggest that multiple kinases may phosphorylate FRQ in vivo.

Original languageEnglish (US)
Pages (from-to)41064-41072
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number44
DOIs
StatePublished - Nov 2 2001

Fingerprint

Neurospora
Calcium-Calmodulin-Dependent Protein Kinases
Circadian Clocks
Clocks
Phosphotransferases
Phosphorylation
Proteins
Calmodulin
Germination
Circadian Rhythm
Cell Extracts
Growth and Development
Redundancy
Genes
Calcium
Phenotype
Light

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of a Calcium/Calmodulin-dependent Protein Kinase That Phosphorylates the Neurospora Circadian Clock Protein FREQUENCY. / Yang, Yuhong; Cheng, Ping; Zhi, Gang; Liu, Yi.

In: Journal of Biological Chemistry, Vol. 276, No. 44, 02.11.2001, p. 41064-41072.

Research output: Contribution to journalArticle

@article{fe338472d2074456a203b5f8c950d432,
title = "Identification of a Calcium/Calmodulin-dependent Protein Kinase That Phosphorylates the Neurospora Circadian Clock Protein FREQUENCY",
abstract = "Phosphorylation of circadian clock proteins represents a major regulatory step that controls circadian clocks. In Neurospora, the circadian clock protein FREQUENCY (FRQ) is progressively phosphorylated over time, and its level decreases when it is hyperphosphorylated. In this study, we showed that most of the kinase activity phosphorylating FRQ in vitro was calcium/ calmodulin-dependent, and the endogenous FRQ in the Neurospora extracts was phosphorylated by a Ca/ CaM-dependent kinase-like activity. From Neurospora cell extracts, an ∼50-kDa Ca/CaM-dependent kinase (CAMK-1) that can specifically phosphorylate FRQ was purified. In vitro, this kinase accounts for near half of the FRQ kinase activity, and it can phosphorylate the FRQ region that contains the three known functionally important phosphorylation sites. To understand the function of camk-I in vivo, it was disrupted in Neurospora by gene replacement. After germination from ascospores, the camk-1 null strains grew slowly, indicating that CAMK-1 plays an important role in growth and development of Neurospora. This phenotype was transient however, revealing redundancy in the system. Analysis of the camk-1 null strain revealed that the deletion of camk-1 affected phase, period, and light-induced phase shifting of the circadian conidiation rhythm. Taken together, our results suggest that multiple kinases may phosphorylate FRQ in vivo.",
author = "Yuhong Yang and Ping Cheng and Gang Zhi and Yi Liu",
year = "2001",
month = "11",
day = "2",
doi = "10.1074/jbc.M106905200",
language = "English (US)",
volume = "276",
pages = "41064--41072",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "44",

}

TY - JOUR

T1 - Identification of a Calcium/Calmodulin-dependent Protein Kinase That Phosphorylates the Neurospora Circadian Clock Protein FREQUENCY

AU - Yang, Yuhong

AU - Cheng, Ping

AU - Zhi, Gang

AU - Liu, Yi

PY - 2001/11/2

Y1 - 2001/11/2

N2 - Phosphorylation of circadian clock proteins represents a major regulatory step that controls circadian clocks. In Neurospora, the circadian clock protein FREQUENCY (FRQ) is progressively phosphorylated over time, and its level decreases when it is hyperphosphorylated. In this study, we showed that most of the kinase activity phosphorylating FRQ in vitro was calcium/ calmodulin-dependent, and the endogenous FRQ in the Neurospora extracts was phosphorylated by a Ca/ CaM-dependent kinase-like activity. From Neurospora cell extracts, an ∼50-kDa Ca/CaM-dependent kinase (CAMK-1) that can specifically phosphorylate FRQ was purified. In vitro, this kinase accounts for near half of the FRQ kinase activity, and it can phosphorylate the FRQ region that contains the three known functionally important phosphorylation sites. To understand the function of camk-I in vivo, it was disrupted in Neurospora by gene replacement. After germination from ascospores, the camk-1 null strains grew slowly, indicating that CAMK-1 plays an important role in growth and development of Neurospora. This phenotype was transient however, revealing redundancy in the system. Analysis of the camk-1 null strain revealed that the deletion of camk-1 affected phase, period, and light-induced phase shifting of the circadian conidiation rhythm. Taken together, our results suggest that multiple kinases may phosphorylate FRQ in vivo.

AB - Phosphorylation of circadian clock proteins represents a major regulatory step that controls circadian clocks. In Neurospora, the circadian clock protein FREQUENCY (FRQ) is progressively phosphorylated over time, and its level decreases when it is hyperphosphorylated. In this study, we showed that most of the kinase activity phosphorylating FRQ in vitro was calcium/ calmodulin-dependent, and the endogenous FRQ in the Neurospora extracts was phosphorylated by a Ca/ CaM-dependent kinase-like activity. From Neurospora cell extracts, an ∼50-kDa Ca/CaM-dependent kinase (CAMK-1) that can specifically phosphorylate FRQ was purified. In vitro, this kinase accounts for near half of the FRQ kinase activity, and it can phosphorylate the FRQ region that contains the three known functionally important phosphorylation sites. To understand the function of camk-I in vivo, it was disrupted in Neurospora by gene replacement. After germination from ascospores, the camk-1 null strains grew slowly, indicating that CAMK-1 plays an important role in growth and development of Neurospora. This phenotype was transient however, revealing redundancy in the system. Analysis of the camk-1 null strain revealed that the deletion of camk-1 affected phase, period, and light-induced phase shifting of the circadian conidiation rhythm. Taken together, our results suggest that multiple kinases may phosphorylate FRQ in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0035798639&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035798639&partnerID=8YFLogxK

U2 - 10.1074/jbc.M106905200

DO - 10.1074/jbc.M106905200

M3 - Article

C2 - 11551951

AN - SCOPUS:0035798639

VL - 276

SP - 41064

EP - 41072

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 44

ER -