C9orf72/ALFA-1 controls TFEB/HLH-30-dependent metabolism through dynamic regulation ofrag GTPases.

The objective of this experiment was to determine changes in gene expression upon loss of alfa-1 in C.elegans under well-fed and starvation conditions. The human C9orf72 has an orthologue in C. elegans based on sequence homology, which is the F18A1.6 gene and also recently named as alfa-1 (ALS/FTD-associated gene homolog 1). We analyzed the domain structure of ALFA-1 and found that it shares the same DENN domains as human C9orf72, including the uDENN, cDENN, and dDENN domains. We obtained a mutant strain of the alfa-1 gene, which has a mutant allele (ok3062) of 486 bp deletion and 24 bp insertion in the region of exon 3 and exon 4. To understand the changes in gene expression in the alfa-1 mutant C. elegans, we performed the microarray gene expression profiling experiments under the defined conditions.本实验旨在探究秀丽隐杆线虫（C. elegans）在正常喂食与饥饿培养条件下，alfa-1基因缺失后其基因表达的变化情况。基于序列同源性分析，人类C9orf72基因在秀丽隐杆线虫中存在同源基因F18A1.6，该基因近期亦被命名为alfa-1（ALS/FTD相关基因同源物1）。我们对ALFA-1的蛋白质结构域进行了分析，发现其与人类C9orf72拥有相同的DENN结构域，包括uDENN、cDENN及dDENN结构域。我们获取了alfa-1基因的突变株，该突变株携带突变等位基因ok3062，其在第3外显子与第4外显子区域存在486 bp的缺失与24 bp的插入。为探明alfa-1突变型秀丽隐杆线虫的基因表达变化，我们在预设实验条件下开展了基因芯片（microarray）表达谱分析实验。