Gimeno C J, Fink G R
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge 02142.
Mol Cell Biol. 1994 Mar;14(3):2100-12. doi: 10.1128/mcb.14.3.2100-2112.1994.
When starved for nitrogen, MATa/MAT alpha cells of the budding yeast Saccharomyces cerevisiae undergo a dimorphic transition to pseudohyphal growth. A visual genetic screen, called PHD (pseudohyphal determinant), for S. cerevisiae pseudohyphal growth mutants was developed. The PHD screen was used to identify seven S. cerevisiae genes that when overexpressed in MATa/MAT alpha cells growing on nitrogen starvation medium cause precocious and unusually vigorous pseudohyphal growth. PHD1, a gene whose overexpression induced invasive pseudohyphal growth on a nutritionally rich medium, was characterized. PHD1 maps to chromosome XI and is predicted to encode a 366-amino-acid protein. PHD1 has a SWI4- and MBP1-like DNA binding motif that is 73% identical over 100 amino acids to a region of Aspergillus nidulans StuA. StuA regulates two pseudohyphal growth-like cell divisions during conidiophore morphogenesis. Epitope-tagged PHD1 was localized to the nucleus by indirect immunofluorescence. These facts suggest that PHD1 may function as a transcriptional regulatory protein. Overexpression of PHD1 in wild-type haploid strains does not induce pseudohyphal growth. Interestingly, PHD1 overexpression enhances pseudohyphal growth in a haploid strain that has the diploid polar budding pattern because of a mutation in the BUD4 gene. In addition, wild-type diploid strains lacking PHD1 undergo pseudohyphal growth when starved for nitrogen. The possible functions of PHD1 in pseudohyphal growth and the uses of the PHD screen to identify morphogenetic regulatory genes from heterologous organisms are discussed.
当处于氮饥饿状态时,出芽酵母酿酒酵母的 MATa/MATα 细胞会经历双态转变,形成假菌丝生长。开发了一种名为 PHD(假菌丝决定因子)的视觉遗传筛选方法,用于筛选酿酒酵母假菌丝生长突变体。PHD 筛选用于鉴定七个酿酒酵母基因,这些基因在氮饥饿培养基上生长的 MATa/MATα 细胞中过表达时,会导致早熟且异常旺盛的假菌丝生长。对 PHD1 进行了表征,该基因的过表达会在营养丰富的培养基上诱导侵袭性假菌丝生长。PHD1 定位于第十一条染色体,预计编码一个 366 个氨基酸的蛋白质。PHD1 具有与 SWI4 和 MBP1 相似的 DNA 结合基序,在 100 个氨基酸上与构巢曲霉 StuA 的一个区域有 73% 的同一性。StuA 在分生孢子梗形态发生过程中调节两个类似假菌丝生长的细胞分裂。通过间接免疫荧光将表位标记的 PHD1 定位到细胞核。这些事实表明 PHD1 可能作为一种转录调节蛋白发挥作用。在野生型单倍体菌株中过表达 PHDl 不会诱导假菌丝生长。有趣的是,由于 BUD4 基因突变,PHD1 过表达增强了具有二倍体极性出芽模式的单倍体菌株中的假菌丝生长。此外,缺乏 PHD1 的野生型二倍体菌株在氮饥饿时会经历假菌丝生长。讨论了 PHD1 在假菌丝生长中的可能功能以及 PHD 筛选用于从异源生物体中鉴定形态发生调节基因的用途。