Yang Qin, Kempken Frank
Department of Genetics and Molecular Biology, Botanical Institute and Botanic Garden, Kiel University, Kiel, Germany.
Front Microbiol. 2020 Sep 18;11:2115. doi: 10.3389/fmicb.2020.02115. eCollection 2020.
MCC/eisosomes are protein-organized domains in the plasma membrane of fungi and algae. However, the composition and function(s) of MCC/eisosomes in the filamentous fungus were previously unknown. To identify proteins that localize to MCC/eisosomes in , we isolated proteins that co-purified with the core MCC/eisosome protein LSP-1, which was tagged with GFP. Proteins that co-fractionated with LSP-1:GFP were then identified by mass spectrometry. Eighteen proteins were GFP-tagged and used to identify six proteins that highly colocalized with the MCC/eisosome marker LSP-1:RFP, while five other proteins showed partial overlap with MCC/eisosomes. Seven of these proteins showed amino acid sequence homology with proteins known to localize to MCC/eisosomes in the yeast . However, homologs of three proteins known to localize to MCC/eisosomes in (Can1, Pkh1/2, and Fhn1) were not found to colocalize with MCC/eisosome proteins in by fluorescence microscopy. Interestingly, one new eisosome protein (glutamine-fructose-6-phosphate aminotransferase, gene ID: NCU07366) was detected in our studies. These findings demonstrate that there are interspecies differences of the protein composition of MCC/eisosomes. To gain further insight, molecular modeling and bioinformatics analysis of the identified proteins were used to propose the organization of MCC/eisosomes in . A model will be discussed for how the broad range of functions predicted for the proteins localized to MCC/eisosomes, including cell wall synthesis, response and signaling, transmembrane transport, and actin organization, suggests that MCC/eisosomes act as organizing centers in the plasma membrane.
MCC/内质体小体是真菌和藻类质膜中由蛋白质组织形成的结构域。然而,丝状真菌中MCC/内质体小体的组成和功能此前尚不清楚。为了鉴定定位于丝状真菌中MCC/内质体小体的蛋白质,我们分离了与用绿色荧光蛋白(GFP)标记的核心MCC/内质体小体蛋白LSP-1共纯化的蛋白质。然后通过质谱鉴定与LSP-1:GFP共分离的蛋白质。18种蛋白质被标记上绿色荧光蛋白,并用于鉴定与MCC/内质体小体标记物LSP-1:红色荧光蛋白(RFP)高度共定位的6种蛋白质,而其他5种蛋白质与MCC/内质体小体有部分重叠。这些蛋白质中有7种与已知定位于酵母中MCC/内质体小体的蛋白质具有氨基酸序列同源性。然而,通过荧光显微镜未发现已知定位于酵母中MCC/内质体小体的3种蛋白质(Can1、Pkh1/2和Fhn1)的同源物与丝状真菌中的MCC/内质体小体蛋白共定位。有趣的是,在我们的研究中检测到一种新的内质体小体蛋白(谷氨酰胺-果糖-6-磷酸转氨酶,基因ID:NCU07366)。这些发现表明MCC/内质体小体的蛋白质组成存在种间差异。为了进一步深入了解,我们对鉴定出的蛋白质进行了分子建模和生物信息学分析,以推测丝状真菌中MCC/内质体小体的结构。将讨论一个模型,该模型表明定位于MCC/内质体小体的蛋白质所具有的广泛功能,包括细胞壁合成、应答和信号传导、跨膜运输以及肌动蛋白组织,提示MCC/内质体小体在质膜中充当组织中心。