Phosphorylation‐dependent interactions between enzymes of plant metabolism and 14‐3‐3 proteins

Moorhead, Greg B. G.; Douglas, Pauline; Cotelle, Valérie; Harthill, J E; Morrice, Nick; Meek, Sarah; Deiting, Uta; Stitt, Mark; Scarabel, Marie; Aitken, Alastair; MacKintosh, Carol

doi:10.1046/j.1365-313x.1999.00417.x

Cited by 266 publications

(250 citation statements)

References 40 publications

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“…The fact that in most F o F 1 isolation procedures no precautions are taken to inhibit phosphatase activity (4,25,35) may offer an explanation why, thus far, 14-3-3 was not identified as a regulatory protein of the ATP synthases. Our findings are in line with a recent report that the chloroplast F 1 ␤-subunit from cauliflower binds to a yeast 14-3-3 affinity column (33). We also demonstrated that purified MF o F 1 and CF o F 1 complexes bind to 14-3-3B affinity columns and are specifically eluted with phosphorylated NR-peptide (data not shown).…”

Section: Discussionsupporting

confidence: 93%

“…In mammalian and yeast cells, many targets for 14-3-3 proteins can be classified as signaling proteins (45), cell cycle regulators (46), and proteins involved in apoptosis (47), whereas plant 14-3-3 targets are notably involved in primary metabolism (19,33). Examples of the latter are the plasma membrane H ϩ ATPase, NR, SPS, trehalose-6-phosphate synthase, and also, as shown here, the primary ATP producers of the cell.…”

Section: Discussionmentioning

confidence: 99%

“…The fact that the uncoupler S13 still doubled the rate of oxygen uptake indicates that the effect of 14-3-3 is caused by blocking the ATP synthase rather than affecting complexes III and IV in the inner mitochondrial membrane. Phosphoserine peptides that match the binding sequences in 14-3-3 target proteins have been used successfully to competitively inhibit the binding of 14-3-3 proteins (18,33,34). We synthesized a phosphopeptide matching the 14-3-3 interaction domain in barley NR (18) containing a phosphorylated (*) motif (RSTS*TP) identical to the one found in Raf-kinase (34).…”

Section: -3-3 Isoforms Are Present Within Plant Mitochondriamentioning

confidence: 99%

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14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase

Bunney

Walraven

Boer

2001

Proc. Natl. Acad. Sci. U.S.A.

182

124

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Mitochondrial and chloroplast ATP synthases are key enzymes in plant metabolism, providing cells with ATP, the universal energy currency. ATP synthases use a transmembrane electrochemical proton gradient to drive synthesis of ATP. The enzyme complexes function as miniature rotary engines, ensuring energy coupling with very high efficiency. Although our understanding of the structure and functioning of the synthase has made enormous progress in recent years, our understanding of regulatory mechanisms is still rather preliminary. Here we report a role for 14-3-3 proteins in the regulation of ATP synthases. These 14-3-3 proteins are highly conserved phosphoserine͞phosphothreonine-binding proteins that regulate a wide range of enzymes in plants, animals, and yeast. Recently, the presence of 14-3-3 proteins in chloroplasts was illustrated, and we show here that plant mitochondria harbor 14-3-3s within the inner mitochondrial-membrane compartment. There, the 14-3-3 proteins were found to be associated with the ATP synthases, in a phosphorylation-dependent manner, through direct interaction with the F 1 ␤-subunit. The activity of the ATP synthases in both organelles is drastically reduced by recombinant 14-3-3. The rapid reduction in chloroplast ATPase activity during dark adaptation was prevented by a phosphopeptide containing the 14-3-3 interaction motif, demonstrating a role for endogenous 14-3-3 in the down-regulation of the CF oF1 activity. We conclude that regulation of the ATP synthases by 14-3-3 represents a mechanism for plant adaptation to environmental changes such as light͞dark transitions, anoxia in roots, and fluctuations in nutrient supply.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: -3-3 Isoforms Are Present Within Plant Mitochondriamentioning

confidence: 99%

See 1 more Smart Citation

14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase

Bunney

Walraven

Boer

2001

Proc. Natl. Acad. Sci. U.S.A.

182

124

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“…Phosphorylation of GS may be important for its stability as found in Brassica napus (Finnemann and Schjoerring 2000). However, activity of GS was not influenced by 14-3-3 binding (Moorhead et al 1999, Riedel et al 2001. As for NR, 14-3-3 proteins may be important for the regulation of GS; however, this point is still not clarified.…”

Section: Post-translational Regulation Of Gsmentioning

confidence: 97%

Light Regulation of Nitrate uptake, Assimilation and Metabolism

Lillo

2004

Plant Ecophysiology

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“…The cells were disrupted by sonication and soluble proteins were then separated by centrifugation (36 900g for 30 minutes at 4°C). The 14-3-3 fusion protein was purified by a Ni-NTA agarose column (Qiagen, Valencia, CA), 19,20 and after overnight thrombin (GE Healthcare, Little Chalfont, United Kingdom) digestion at 23°C the 14-3-3 part was eluted. The sample was further purified by HiTrap Q ion exchange chromatography (Amersham Biosciences) in 20 mM Tris-HCl, pH 8.0, with NaCl gradient elution.…”

Section: Protein Production and Purificationmentioning

confidence: 99%

β2 integrin phosphorylation on Thr758 acts as a molecular switch to regulate 14-3-3 and filamin binding

Takala

Nurminen

Nurmi³

et al. 2008

Blood

149

235

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Leukocyte integrins of the ␤2 family are essential for immune cell-cell adhesion. In activated cells, ␤2 integrins are phosphorylated on the cytoplasmic Thr758, leading to 14-3-3 protein recruitment to the ␤2 integrin. The mutation of this phosphorylation site impairs cell adhesion, actin reorganization, and cell spreading. Thr758 is contained in a Thr triplet of ␤2 that also mediates binding to filamin. Here, we investigated the binding of filamin, talin, and 14-3-3 proteins to phosphorylated and unphosphorylated ␤2 integrins by biochemical methods and x-ray crystallography. 14-3-3 proteins bound only to the phosphorylated integrin cytoplasmic peptide, with a high affinity (K d , 261 nM), whereas filamin bound only the unphosphorylated integrin cytoplasmic peptide (K d , 0.5 mM). Phosphorylation did not regulate talin binding to ␤2 directly, but 14-3-3 was able to outcompete talin for the binding to phosphorylated ␤2 integrin. X-ray crystallographic data clearly IntroductionIntegrins are heterodimeric plasma membrane receptors that mediate binding to the extracellular matrix and to ligands present on the surface of other cells. Their function is tightly regulated; they bind ligands only after activation. Modulation of integrin activity occurs through tightly regulated interactions between cytoplasmic molecules and integrin intracellular tails. Factors binding to integrin cytoplasmic domains regulating integrin adhesiveness include the cytoskeletal proteins talin 1,2 and filamin, 3 and the 14-3-3 proteins, which are molecular adaptors that bind to phosphorylated serine or threonine (pSer/ pThr) containing polypeptide sequences. 4 The ␤2 integrins are expressed exclusively on leukocytes and bind ICAM molecules on other leukocytes and endothelial cells after cell activation. 5,6 Talin binds to ␤2 integrins in vitro and in cells and is involved in activating the ␤2 integrins, resulting in binding to ICAMs. 1,4,[7][8][9] The ␤2 integrin polypeptide chain is phosphorylated on the intracellular domain on several residues after cell stimulation with various agents. 10 Thr758 is a physiologically important amino acid residue in the ␤2 cytoplasmic tail, and becomes phosphorylated after T-cell stimulation with T-cell receptor (TCR) antibodies or with phorbol esters. [11][12][13] After its phosphorylation, ␤2 binds to 14-3-3 proteins both in vitro and in cells. 4 Blocking of this interaction with a ␤2 Thr758 to Ala mutation, or by expression of constructs that bind to 14-3-3 proteins and block their interactions with target proteins, leads to abrogation of actin cytoskeleton rearrangements, cell spreading, and adhesion to ICAM ligands. 4 ␤2-Thr758 phosphorylation leads to the activation of the actin cytoskeleton modulators, Rac1/Cdc42, in cells. 13 The region in the ␤2 cytoplasmic tail that binds 14-3-3 proteins has been reported to interact with filamin in other integrins, 14 and for the strong filamin-binder ␤7 integrin, phosphorylation mimicking substitutions of 3 threonine residues (TTT) reduces filamin affinity. 3 Fi...

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Phosphorylation‐dependent interactions between enzymes of plant metabolism and 14‐3‐3 proteins

Cited by 266 publications

References 40 publications

14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase

14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase

Light Regulation of Nitrate uptake, Assimilation and Metabolism

β2 integrin phosphorylation on Thr758 acts as a molecular switch to regulate 14-3-3 and filamin binding

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