Isozyme-specific modes of activation of CTP: Phosphorylcholine cytidylyltransferase in arabidopsis thaliana at low temperature

Rie Inatsugi; Hiromitsu Kawai; Yasuyo Yamaoka; Yanbo Yu; Akira Sekiguchi; Masanobu Nakamura; Ikuo Nishida

doi:10.1093/pcp/pcp115

Isozyme-specific modes of activation of CTP: Phosphorylcholine cytidylyltransferase in arabidopsis thaliana at low temperature

Rie Inatsugi, Hiromitsu Kawai, Yasuyo Yamaoka, Yanbo Yu, Akira Sekiguchi, Masanobu Nakamura, Ikuo Nishida

Biotechnology

Research output: Contribution to journal › Article › peer-review

31 Scopus citations

Abstract

Arabidopsis thaliana increases cellular phosphatidylcholine (PC) content during cold acclimation by up-regulating PC biosynthesis. The A. thaliana genes CCT1 and CCT2 encode CTP:phosphorylcholine cytidylyltransferases (CCTs; EC 2.7.7.15), which regulate PC biosynthesis via the CDPcholine pathway. We isolated the T-DNA-tagged knockout mutants cct1 and cct2 of A. thaliana (Wassilevskaja; WS). CCT activity in cct1 and cct2 plants accounted for 29 and 79 to the cellular CCT activity of WS plants, respectively. When plants were exposed to 2°C for 7 d, CCT activity increased in both cct1 and cct2 plants, and immunoblot analyses revealed that cct1 contained an increased level of CCT2 protein whereas cct2 exhibited little increase in CCT1 level. For each mutant grown at 23°C, CCT activity was mainly enriched in the particulate (15,000 × g pellet) and microsomal (150,000 × g pellet) fractions from rosette leaf homogenates. After exposure to cold, the particulate and microsomal fractions of cct1 plants had higher total CCT activity due to increased levels of CCT2; in contrast, the levels of CCT1 in cct2 plants remained unchanged in particulate and microsomal fractions despite a significant increase in the total CCT activity. We conclude that the CDPcholine pathway of A. thaliana is up-regulated at low temperature via isogene-specific modes: enhanced expression of CCT2 and post-translational activation/inactivation of CCT1 in membranes. PC levels were similarly maintained in both mutants and WS plants after 14 d at 2°C, suggesting that either of the CCT genes is sufficient for PC biosynthesis at low temperature.

Original language	English
Pages (from-to)	1727-1735
Number of pages	9
Journal	Plant and Cell Physiology
Volume	50
Issue number	10
DOIs	https://doi.org/10.1093/pcp/pcp115
State	Published - Oct 2009

Bibliographical note

Funding Information:
The Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN); the Ministry of Education, Culture, Sports, Science, and Technology (Grant-in-Aid for Priority Areas 17051004); the Japanese Society for the Promotion of Science (Grant-in-Aid for Scientific Research (C), 21570034).

Keywords

Arabidopsis thaliana
CDPcholine pathway
Cold acclimation
Membrane expansion
Phosphati-dylcholine
Post-translational regulation

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title = "Isozyme-specific modes of activation of CTP: Phosphorylcholine cytidylyltransferase in arabidopsis thaliana at low temperature",

abstract = "Arabidopsis thaliana increases cellular phosphatidylcholine (PC) content during cold acclimation by up-regulating PC biosynthesis. The A. thaliana genes CCT1 and CCT2 encode CTP:phosphorylcholine cytidylyltransferases (CCTs; EC 2.7.7.15), which regulate PC biosynthesis via the CDPcholine pathway. We isolated the T-DNA-tagged knockout mutants cct1 and cct2 of A. thaliana (Wassilevskaja; WS). CCT activity in cct1 and cct2 plants accounted for 29 and 79 to the cellular CCT activity of WS plants, respectively. When plants were exposed to 2°C for 7 d, CCT activity increased in both cct1 and cct2 plants, and immunoblot analyses revealed that cct1 contained an increased level of CCT2 protein whereas cct2 exhibited little increase in CCT1 level. For each mutant grown at 23°C, CCT activity was mainly enriched in the particulate (15,000 × g pellet) and microsomal (150,000 × g pellet) fractions from rosette leaf homogenates. After exposure to cold, the particulate and microsomal fractions of cct1 plants had higher total CCT activity due to increased levels of CCT2; in contrast, the levels of CCT1 in cct2 plants remained unchanged in particulate and microsomal fractions despite a significant increase in the total CCT activity. We conclude that the CDPcholine pathway of A. thaliana is up-regulated at low temperature via isogene-specific modes: enhanced expression of CCT2 and post-translational activation/inactivation of CCT1 in membranes. PC levels were similarly maintained in both mutants and WS plants after 14 d at 2°C, suggesting that either of the CCT genes is sufficient for PC biosynthesis at low temperature.",

keywords = "Arabidopsis thaliana, CDPcholine pathway, Cold acclimation, Membrane expansion, Phosphati-dylcholine, Post-translational regulation",

author = "Rie Inatsugi and Hiromitsu Kawai and Yasuyo Yamaoka and Yanbo Yu and Akira Sekiguchi and Masanobu Nakamura and Ikuo Nishida",

note = "Funding Information: The Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN); the Ministry of Education, Culture, Sports, Science, and Technology (Grant-in-Aid for Priority Areas 17051004); the Japanese Society for the Promotion of Science (Grant-in-Aid for Scientific Research (C), 21570034).",

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doi = "10.1093/pcp/pcp115",

language = "English",

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pages = "1727--1735",

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T2 - Phosphorylcholine cytidylyltransferase in arabidopsis thaliana at low temperature

AU - Inatsugi, Rie

AU - Kawai, Hiromitsu

AU - Yamaoka, Yasuyo

AU - Yu, Yanbo

AU - Sekiguchi, Akira

AU - Nakamura, Masanobu

AU - Nishida, Ikuo

N1 - Funding Information: The Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN); the Ministry of Education, Culture, Sports, Science, and Technology (Grant-in-Aid for Priority Areas 17051004); the Japanese Society for the Promotion of Science (Grant-in-Aid for Scientific Research (C), 21570034).

PY - 2009/10

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N2 - Arabidopsis thaliana increases cellular phosphatidylcholine (PC) content during cold acclimation by up-regulating PC biosynthesis. The A. thaliana genes CCT1 and CCT2 encode CTP:phosphorylcholine cytidylyltransferases (CCTs; EC 2.7.7.15), which regulate PC biosynthesis via the CDPcholine pathway. We isolated the T-DNA-tagged knockout mutants cct1 and cct2 of A. thaliana (Wassilevskaja; WS). CCT activity in cct1 and cct2 plants accounted for 29 and 79 to the cellular CCT activity of WS plants, respectively. When plants were exposed to 2°C for 7 d, CCT activity increased in both cct1 and cct2 plants, and immunoblot analyses revealed that cct1 contained an increased level of CCT2 protein whereas cct2 exhibited little increase in CCT1 level. For each mutant grown at 23°C, CCT activity was mainly enriched in the particulate (15,000 × g pellet) and microsomal (150,000 × g pellet) fractions from rosette leaf homogenates. After exposure to cold, the particulate and microsomal fractions of cct1 plants had higher total CCT activity due to increased levels of CCT2; in contrast, the levels of CCT1 in cct2 plants remained unchanged in particulate and microsomal fractions despite a significant increase in the total CCT activity. We conclude that the CDPcholine pathway of A. thaliana is up-regulated at low temperature via isogene-specific modes: enhanced expression of CCT2 and post-translational activation/inactivation of CCT1 in membranes. PC levels were similarly maintained in both mutants and WS plants after 14 d at 2°C, suggesting that either of the CCT genes is sufficient for PC biosynthesis at low temperature.

AB - Arabidopsis thaliana increases cellular phosphatidylcholine (PC) content during cold acclimation by up-regulating PC biosynthesis. The A. thaliana genes CCT1 and CCT2 encode CTP:phosphorylcholine cytidylyltransferases (CCTs; EC 2.7.7.15), which regulate PC biosynthesis via the CDPcholine pathway. We isolated the T-DNA-tagged knockout mutants cct1 and cct2 of A. thaliana (Wassilevskaja; WS). CCT activity in cct1 and cct2 plants accounted for 29 and 79 to the cellular CCT activity of WS plants, respectively. When plants were exposed to 2°C for 7 d, CCT activity increased in both cct1 and cct2 plants, and immunoblot analyses revealed that cct1 contained an increased level of CCT2 protein whereas cct2 exhibited little increase in CCT1 level. For each mutant grown at 23°C, CCT activity was mainly enriched in the particulate (15,000 × g pellet) and microsomal (150,000 × g pellet) fractions from rosette leaf homogenates. After exposure to cold, the particulate and microsomal fractions of cct1 plants had higher total CCT activity due to increased levels of CCT2; in contrast, the levels of CCT1 in cct2 plants remained unchanged in particulate and microsomal fractions despite a significant increase in the total CCT activity. We conclude that the CDPcholine pathway of A. thaliana is up-regulated at low temperature via isogene-specific modes: enhanced expression of CCT2 and post-translational activation/inactivation of CCT1 in membranes. PC levels were similarly maintained in both mutants and WS plants after 14 d at 2°C, suggesting that either of the CCT genes is sufficient for PC biosynthesis at low temperature.

KW - Arabidopsis thaliana

KW - CDPcholine pathway

KW - Cold acclimation

KW - Membrane expansion

KW - Phosphati-dylcholine

KW - Post-translational regulation

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DO - 10.1093/pcp/pcp115

M3 - Article

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AN - SCOPUS:70349847645

SN - 0032-0781

VL - 50

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EP - 1735

JO - Plant and Cell Physiology

JF - Plant and Cell Physiology

IS - 10

ER -

Isozyme-specific modes of activation of CTP: Phosphorylcholine cytidylyltransferase in arabidopsis thaliana at low temperature

Abstract

Bibliographical note

Keywords

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