Saupe S J, Kuldau G A, Smith M L, Glass N L
Botany Department, University of British Columbia, Vancouver, Canada.
Genetics. 1996 Aug;143(4):1589-600. doi: 10.1093/genetics/143.4.1589.
Filamentous fungi are capable of hyphal fusion, but heterokaryon formation between different isolates is controlled by specific loci termed het loci. Heterokaryotic cells formed between strains of different het genotype are rapidly destroyed or strongly inhibited in their growth. In Neurospora crassa, at least 11 loci, including the mating type locus, affect the capacity to form a heterokaryon between different isolates. In this report, we describe the molecular characterization of the vegetative incompatibility locus, het-C. The het-COR allele was cloned by genetically identifying the het-C locus in a chromosome walk, and the activity of clones containing the het-COR allele was tested in a functional transformation assay. The het-COR allele encodes a 966-amino acid polypeptide with a putative signal peptide, a coiled-coil motif and a C-terminal glycine-rich domain, similar to glycine-rich domains detected in various extracellular and structural cell envelope proteins. Both the coiled-coil and one-third of the glycine-rich carboxyl terminal domains were required for full het-COR activity. Mutants of het-COR were obtained by repeat-induced point mutation (RIP); these mutants were indistinguishable from wild type during vegetative growth and sexual reproduction but displayed dual compatibility with both of two mutually incompatible het-COR and het-cPA strains.
丝状真菌能够进行菌丝融合,但不同分离株之间异核体的形成受特定基因座(称为het基因座)控制。在不同het基因型的菌株之间形成的异核体细胞会迅速被破坏或其生长受到强烈抑制。在粗糙脉孢菌中,至少有11个基因座,包括交配型基因座,影响不同分离株之间形成异核体的能力。在本报告中,我们描述了营养不亲和基因座het-C的分子特征。通过染色体步移法对het-C基因座进行遗传鉴定,克隆了het-COR等位基因,并在功能转化试验中测试了含有het-COR等位基因的克隆的活性。het-COR等位基因编码一个含有推定信号肽、卷曲螺旋基序和C端富含甘氨酸结构域的966个氨基酸的多肽,类似于在各种细胞外和结构细胞包膜蛋白中检测到的富含甘氨酸的结构域。卷曲螺旋结构域和富含甘氨酸的羧基末端结构域的三分之一对于het-COR的完全活性都是必需的。通过重复诱导点突变(RIP)获得了het-COR的突变体;这些突变体在营养生长和有性生殖过程中与野生型没有区别,但与两个相互不亲和的het-COR和het-cPA菌株都表现出双重相容性。