Hoecker Natalie, Honke Anna, Frey Katharina, Leister Dario, Schneider Anja
Molekularbiologie der Pflanze (Botanik), Department Biologie I, Ludwig-Maximilians-Universität München, 82152 Martinsried, Germany.
Plants (Basel). 2020 Feb 13;9(2):239. doi: 10.3390/plants9020239.
Chloroplast manganese transporter 1 (CMT1) and photosynthesis-affected mutant 71 (PAM71) are two membrane proteins that function sequentially to mediate the passage of manganese across the chloroplast envelope and the thylakoid membrane. CMT1 and PAM71 belong to a small five-member protein family in . The other three, photosynthesis-affected mutant 71 like 3 (PML3), PML4 and PML5 are not predicted to reside in chloroplast membranes. In this study, the subcellular localization of PML3:GFP, PML4:GFP and PML5:GFP was determined using transient and stable expression assays. PML3:GFP localizes to the Golgi apparatus, whereas PML4:GFP and PML5:GFP are found in the endoplasmic reticulum. We also examined patterns of , and promoter activity. Although the precise expression pattern of each promoter was unique, all three genes were expressed in the leaf vasculature and in roots. Greenhouse grown single mutants , , and the double mutant did not exhibit growth defects, however an inspection of the root growth revealed a difference between and the other genotypes, including wild-type, in 500 µM manganese growth conditions. Strikingly, overexpression of PML3 resulted in a stunted growth phenotype. Putative functions of PML3, PML4 and PML5 are discussed in light of what is known about PAM71 and CMT1.
叶绿体锰转运蛋白1(CMT1)和光合作用受影响突变体71(PAM71)是两种膜蛋白,它们依次发挥作用,介导锰穿过叶绿体包膜和类囊体膜。CMT1和PAM71属于一个包含五个成员的小蛋白家族。另外三个,即类光合作用受影响突变体71样蛋白3(PML3)、PML4和PML5预计并不定位于叶绿体膜。在本研究中,利用瞬时表达和稳定表达分析确定了PML3:GFP、PML4:GFP和PML5:GFP的亚细胞定位。PML3:GFP定位于高尔基体,而PML4:GFP和PML5:GFP在内质网中被发现。我们还检测了 、 和 启动子活性模式。尽管每个启动子的确切表达模式都是独特的,但这三个基因均在叶脉和根中表达。温室种植的单突变体 、 、 以及 双突变体均未表现出生长缺陷,然而在500 μM锰生长条件下对根生长的检查发现, 与包括野生型在内的其他基因型之间存在差异。令人惊讶的是,PML3的过表达导致了生长发育迟缓的表型。结合对PAM71和CMT1的已知信息,讨论了PML3、PML4和PML5的推定功能。