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NAD激酶Slr0400在集胞藻PCC 6803中作为生长抑制因子发挥作用。

The NAD Kinase Slr0400 Functions as a Growth Repressor in Synechocystis sp. PCC 6803.

作者信息

Ishikawa Yuuma, Cassan Cedric, Kadeer Aikeranmu, Yuasa Koki, Sato Nozomu, Sonoike Kintake, Kaneko Yasuko, Miyagi Atsuko, Takahashi Hiroko, Ishikawa Toshiki, Yamaguchi Masatoshi, Nishiyama Yoshitaka, Hihara Yukako, Gibon Yves, Kawai-Yamada Maki

机构信息

Institute for Molecular Physiology and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich-Heine-University Düsseldorf, Düsseldorf 40225, Germany.

Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Chikusa, Nagoya, 464-8601 Japan.

出版信息

Plant Cell Physiol. 2021 Sep 24;62(4):668-677. doi: 10.1093/pcp/pcab023.

Abstract

NADP+, the phosphorylated form of nicotinamide adenine dinucleotide (NAD), plays an essential role in many cellular processes. NAD kinase (NADK), which is conserved in all living organisms, catalyzes the phosphorylation of NAD+ to NADP+. However, the physiological role of phosphorylation of NAD+ to NADP+ in the cyanobacterium Synechocystis remains unclear. In this study, we report that slr0400, an NADK-encoding gene in Synechocystis, functions as a growth repressor under light-activated heterotrophic growth conditions and light and dark cycle conditions in the presence of glucose. We show, via characterization of NAD(P)(H) content and enzyme activity, that NAD+ accumulation in slr0400-deficient mutant results in the unsuppressed activity of glycolysis and tricarboxylic acid (TCA) cycle enzymes. In determining whether Slr0400 functions as a typical NADK, we found that constitutive expression of slr0400 in an Arabidopsis nadk2-mutant background complements the pale-green phenotype. Moreover, to determine the physiological background behind the growth advantage of mutants lacking slr04000, we investigated the photobleaching phenotype of slr0400-deficient mutant under high-light conditions. Photosynthetic analysis found in the slr0400-deficient mutant resulted from malfunctions in the Photosystem II (PSII) photosynthetic machinery. Overall, our results suggest that NADP(H)/NAD(H) maintenance by slr0400 plays a significant role in modulating glycolysis and the TCA cycle to repress the growth rate and maintain the photosynthetic capacity.

摘要

烟酰胺腺嘌呤二核苷酸(NAD)的磷酸化形式NADP⁺在许多细胞过程中起着至关重要的作用。NAD激酶(NADK)在所有生物体中都保守,催化NAD⁺磷酸化为NADP⁺。然而,在蓝藻集胞藻中,NAD⁺磷酸化为NADP⁺的生理作用仍不清楚。在本研究中,我们报道集胞藻中一个编码NADK的基因slr0400在光激活的异养生长条件下以及在有葡萄糖存在的光暗循环条件下起生长抑制因子的作用。通过对NAD(P)(H)含量和酶活性的表征,我们表明slr0400缺陷型突变体中NAD⁺的积累导致糖酵解和三羧酸(TCA)循环酶的活性未受抑制。在确定Slr0400是否作为典型的NADK发挥作用时,我们发现slr0400在拟南芥nadk2突变体背景下的组成型表达弥补了淡绿色表型。此外,为了确定缺乏slr0400的突变体生长优势背后的生理背景,我们研究了slr0400缺陷型突变体在高光条件下的光漂白表型。在slr0400缺陷型突变体中发现的光合分析结果是由于光系统II(PSII)光合机制故障所致。总体而言,我们的结果表明slr0400对NADP(H)/NAD(H)的维持在调节糖酵解和TCA循环以抑制生长速率和维持光合能力方面起着重要作用。

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