微管组织中心(MTOC)是真核细胞中的亚细胞结构,在那里发生微管(MT)的成核,代表细丝的负端。它们的定位取决于物种,细胞类型,细胞周期阶段。沿着真菌王国,细胞核中的纺锤杆体(SPB)(相当于动物细胞中的中心体)是主要的MTOC。在丝状真菌中已鉴定出其他MTOC,例如裂殖酵母菌丝尖端的Spitzenkörper或构巢曲霉的间隔孔。然而,在真菌模型生物中,这些替代MTOC尚未得到认可。这里,我们提供了一个基于质谱的数据集,该数据集与带有荧光蛋白标记的N.crassa的四个MTOC成分相互作用:γ-微管蛋白-sGFP(SPB的主要成核剂),MZT-1-sGFP(结构SPB微蛋白),APS-2-dRFP(间隔蛋白和公认的SPB成分),和SPA-10-sGFP(间隔MTOC蛋白)。包括WT和胞质GFP表达菌株作为对照。通过Co-IP1使用GFP-磁性琼脂糖将蛋白质相互作用体拉低,所述GFP-磁性琼脂糖捕获处于其天然状态的重组GFP蛋白(包括GFP-衍生物)。通过SDS-PAGE分离结合的蛋白质并通过纳米LC-MS/MS2鉴定。使用N.crassa蛋白质数据库进行蛋白质注释。
Microtubule Organizing Centers (
MTOC) are subcellular structures in eukaryotic cells where nucleation of microtubules (MTs) takes place and represents the filament\'s minus end. Their localization depends on the species, cell type, and cell cycle stage. Along the fungal kingdom, the Spindle Pole Body (SPB) in the nucleus (an equivalent to Centrosomes in animal cells) is the principal
MTOC. Other MTOCs have been identified in filamentous fungi, such as the Spitzenkörper in the hyphal tips of Schizosaccharomyces pombe or the septal pore of Aspergillus nidulans. However, in the fungal-model organism Neurospora crassa, these alternative MTOCs have not been recognized. Here, we present a Mass spectrometry-based dataset of proteins interacting with four
MTOC components of N. crassa tagged with fluorescent proteins: γ-Tubulin-sGFP (main nucleator at the SPB), MZT-1-sGFP (structural SPB microprotein), APS-2-dRFP (septal protein and recognized SPB component), and SPA-10-sGFP (septal
MTOC protein). A WT and a cytosolic GFP expressing strain were included as controls. The protein interactors were pulled down by Co-IP1, using GFP-Magnetic agarose that captures recombinant GFP proteins (including GFP-derivatives) in their native state. Bounded proteins were separated by SDS-PAGE and identified by nano LC-MS/MS2. The protein annotation was done using the N. crassa protein database.