Chloroplast envelope ATPase PGA1/AtFtsH12 is required for chloroplast protein accumulation and cytosol-chloroplast protein homeostasis in Arabidopsis.
Chloroplast envelope ATPase PGA1/AtFtsH12 is required for chloroplast protein accumulation and cytosol-chloroplast protein homeostasis in Arabidopsis.
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DOI：
10.1016/j.jbc.2022.102489
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发表时间：
2022-10
影响因子：
4.8
通讯作者：
Qi, Yafei
中科院分区：
文献类型：
--
作者：
Li, Qinglong;Wang, Xiaomin;Lei, Yang;Wang, Yanling;Li, Bilang;Liu, Xiayan;An, Lijun;Yu, Fei;Qi, Yafei
叶绿体发育过程中光合蛋白复合物的建立需要大量由核基因组编码的叶绿体蛋白的涌入，这对细胞质和叶绿体蛋白的稳态和叶绿体发育至关重要。然而，调控这一过程的机制在高等植物中仍未得到很好的理解。在这里，我们报道了浅绿色拟南芥pga1-1突变体的分离和特性，该突变体在叶绿体发育和叶绿体蛋白积累方面存在缺陷。利用遗传和生化证据，我们发现PGA1编码FtsH家族蛋白的叶绿体包膜定位蛋白AtFtsH12。我们确定了保守的atp酶结构域的GAD基序的G703R突变使pga1-1成为必需基因AtFtsH12的一个可行的亚形态等位基因。在去黄化实验中，我们发现pga1-1中光合蛋白的积累和光合基因的表达受到损害。利用FNRctp-GFP和pTAC2-GFP报告基因，我们证明了AtFtsH12是体内叶绿体蛋白积累所必需的。有趣的是，我们发现突变体AtFtsH12基因在pga1-1中的表达增加，这表明存在反馈调节。此外，我们发现pga1-1触发了细胞质和叶绿体的蛋白酶抑制反应。总之，利用新的pga1-1突变体，我们证明了AtFtsH12在叶绿体蛋白稳态和叶绿体发育中的功能。
The establishment of photosynthetic protein complexes during chloroplast development requires the influx of a large number of chloroplast proteins that are encoded by the nuclear genome, which is critical for cytosol and chloroplast protein homeostasis and chloroplast development. However, the mechanisms regulating this process are still not well understood in higher plants. Here, we report the isolation and characterization of the pale green Arabidopsis pga1-1 mutant, which is defective in chloroplast development and chloroplast protein accumulation. Using genetic and biochemical evidence, we reveal that PGA1 encodes AtFtsH12, a chloroplast envelope-localized protein of the FtsH family proteins. We determined a G703R mutation in the GAD motif of the conserved ATPase domain renders the pga1-1 a viable hypomorphic allele of the essential gene AtFtsH12. In de-etiolation assays, we showed that the accumulation of photosynthetic proteins and the expression of photosynthetic genes were impaired in pga1-1. Using the FNRctp-GFP and pTAC2-GFP reporters, we demonstrated that AtFtsH12 was required for the accumulation of chloroplast proteins in vivo. Interestingly, we identified an increase in expression of the mutant AtFtsH12 gene in pga1-1, suggesting a feedback regulation. Moreover, we found that cytosolic and chloroplast proteostasis responses were triggered in pga1-1. Together, taking advantage of the novel pga1-1 mutant, we demonstrate the function of AtFtsH12 in chloroplast protein homeostasis and chloroplast development.
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