Recognition of S-RNases by an S locus F-box like protein and an S haplotype-specific F-box like protein in the Prunus-specific self-incompatibility system
Recognition of S-RNases by an S locus F-box like protein and an S haplotype-specific F-box like protein in the Prunus-specific self-incompatibility system
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DOI：
10.1007/s11103-019-00860-8
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发表时间：
2019-07-01
影响因子：
5.1
通讯作者：
Tao, Ryutaro
中科院分区：
文献类型：
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作者：
Matsumoto, Daiki;Tao, Ryutaro
关键信息在甜樱桃的亲和花粉管中，S-RNase 被证明主要被 S 位点 F 盒样蛋白和 S 单倍型特异性 F 盒样蛋白识别。 摘要 自交不亲和性 (SI) 是一种生殖屏障，它拒绝自体花粉并抑制自体受精以促进异交。在茄科和蔷薇科中，基于 S-RNase 的配子体 SI (GSI) 包含 S-RNase 和 F-box 蛋白，分别作为雌蕊和花粉 S 决定簇。假定相容的花粉管能够解毒内化的细胞毒性 S-RNase 以维持生长。 S-RNase 解毒是由茄科花粉 S 决定簇 S 位点 F-box 蛋白 (SLF) 形成的 Skp1-cullin1-F-box 蛋白复合物 (SCF) 进行的。在李属中，一般抑制剂 (GI)，但不是花粉 S 决定簇 S 单倍型特异性 F-box 蛋白 (SFB)，被假设可以解毒 S-RNase。最近，SLF 样蛋白 1-3 (SLFL1-3) 被建议作为 GI 候选蛋白，尽管其他蛋白仍有可能主要在 GI 中发挥作用。为了鉴定其他 GI 候选者，我们在甜樱桃中分离了其他四种花粉表达的 SLFL 和 SFB 样 (SFBL) 蛋白 PavSLFL6、PavSLFL7A、PavSFBL1 和 PavSFBL2。四种 PavS-RNase 的结合测定表明，PavSFBL2 与 PavS(1, 6)-RNase 结合，而其他则不结合。 PavSFBL2被证实在体外形成SCF复合物。使用重组 PavS(6)-RNase 作为针对花粉提取物的诱饵的免疫共沉淀测定和质谱分析鉴定了 PavSLFL 和 PavSFBL2 的 SCF 复合物成分、M 位点编码的谷胱甘肽 S 转移酶 (MGST)、DnaJ 样蛋白和其他次要蛋白。这些结果表明，SLFL 和 SFBL 可以作为李属特异性 S-RNase 基 GSI 中的主要 GI。
Key messageS-RNase was demonstrated to be predominantly recognized by an S locus F-box-like protein and an S haplotype-specific F-box-like protein in compatible pollen tubes of sweet cherry.AbstractSelf-incompatibility (SI) is a reproductive barrier that rejects self-pollen and inhibits self-fertilization to promote outcrossing. In Solanaceae and Rosaceae, S-RNase-based gametophytic SI (GSI) comprises S-RNase and F-box protein(s) as the pistil and pollen S determinants, respectively. Compatible pollen tubes are assumed to detoxify the internalized cytotoxic S-RNases to maintain growth. S-RNase detoxification is conducted by the Skp1-cullin1-F-box protein complex (SCF) formed by pollen S determinants, S locus F-box proteins (SLFs), in Solanaceae. In Prunus, the general inhibitor (GI), but not pollen S determinant S haplotype-specific F-box protein (SFB), is hypothesized to detoxify S-RNases. Recently, SLF-like proteins 1-3 (SLFL1-3) were suggested as GI candidates, although it is still possible that other proteins function predominantly in GI. To identify the other GI candidates, we isolated four other pollen-expressed SLFL and SFB-like (SFBL) proteins PavSLFL6, PavSLFL7A, PavSFBL1, and PavSFBL2 in sweet cherry. Binding assays with four PavS-RNases indicated that PavSFBL2 bound to PavS(1, 6)-RNase while the others bound to nothing. PavSFBL2 was confirmed to form an SCF complex in vitro. A co-immunoprecipitation assay using the recombinant PavS(6)-RNase as bait against pollen extracts and a mass spectrometry analysis identified the SCF complex components of PavSLFLs and PavSFBL2, M-locus-encoded glutathione S-transferase (MGST), DnaJ-like protein, and other minor proteins. These results suggest that SLFLs and SFBLs could act as predominant GIs in Prunus-specific S-RNase-based GSI.