Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA, 19107, USA.
Laboratory of Cell Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, NY, 10065, USA.
Sci Rep. 2022 Oct 21;12(1):17691. doi: 10.1038/s41598-022-22183-3.
In Saccharomyces cerevisiae, the pre-mRNA leakage 39-kDa protein (ScPml39) was reported to retain unspliced pre-mRNA prior to export through nuclear pore complexes (NPCs). Pml39 homologs outside the Saccharomycetaceae family are currently unknown, and mechanistic insight into Pml39 function is lacking. Here we determined the crystal structure of ScPml39 at 2.5 Å resolution to facilitate the discovery of orthologs beyond Saccharomycetaceae, e.g. in Schizosaccharomyces pombe or human. The crystal structure revealed integrated zf-C3HC and Rsm1 modules, which are tightly associated through a hydrophobic interface to form a single domain. Both zf-C3HC and Rsm1 modules belong to the Zn-containing BIR (Baculovirus IAP repeat)-like super family, with key residues of the canonical BIR domain being conserved. Features unique to the Pml39 modules refer to the spacing between the Zn-coordinating residues, giving rise to a substantially tilted helix αC in the zf-C3HC and Rsm1 modules, and an extra helix αAB' in the Rsm1 module. Conservation of key residues responsible for its distinct features identifies S. pombe Rsm1 and Homo sapiens NIPA/ZC3HC1 as structural orthologs of ScPml39. Based on the recent functional characterization of NIPA/ZC3HC1 as a scaffold protein that stabilizes the nuclear basket of the NPC, our data suggest an analogous function of ScPml39 in S. cerevisiae.
在酿酒酵母中,已报道前 mRNA 渗漏 39kDa 蛋白(ScPml39)在通过核孔复合物(NPC)输出之前保留未剪接的前 mRNA。目前尚不清楚酿酒酵母科以外的 Pml39 同源物,并且对 Pml39 功能的机制见解也缺乏。在这里,我们确定了 ScPml39 的晶体结构,分辨率为 2.5Å,以促进在酿酒酵母科以外的同源物的发现,例如在裂殖酵母或人类中。晶体结构揭示了整合的 zf-C3HC 和 Rsm1 模块,它们通过疏水界面紧密相关,形成单个结构域。zf-C3HC 和 Rsm1 模块都属于含 Zn 的 BIR(杆状病毒 IAP 重复)样超家族,其特征性 BIR 结构域的关键残基被保守。Pml39 模块特有的特征涉及到 Zn 配位残基之间的间隔,导致 zf-C3HC 和 Rsm1 模块中的螺旋 αC 明显倾斜,以及 Rsm1 模块中的额外螺旋 αAB'。负责其独特特征的关键残基的保守性确定裂殖酵母 Rsm1 和人类 NIPA/ZC3HC1 是 ScPml39 的结构同源物。基于最近对 NIPA/ZC3HC1 作为稳定 NPC 核篮的支架蛋白的功能表征,我们的数据表明 ScPml39 在酿酒酵母中具有类似的功能。