P5B ATPases are present in the genomes of diverse unicellular and multicellular eukaryotes, indicating that they have an ancient origin, and that they are important for cellular fitness. Inactivation of ATP13A2, one of the four human P5B ATPases, leads to early-onset Parkinson’s disease (Kufor-Rakeb Syndrome). The presence of an invariant PPALP motif within the putative substrate interaction pocket of transmembrane segment M4 suggests that all P5B ATPases might have similar transport specificity; however, the identity of the transport substrate(s) remains unknown. Nematodes of the genus Caenorhabditis possess three paralogous P5B ATPase genes, catp-5, catp-6 and catp-7, which probably originated from a single ancestral gene around the time of origin of the Caenorhabditid clade. By using CRISPR/Cas9, we have systematically investigated the expression patterns, subcellular localization and biological functions of each of the P5B ATPases of C. elegans. We find that each gene has a unique expression pattern, and that some tissues express more than one P5B. In some tissues where their expression patterns overlap, different P5Bs are targeted to different subcellular compartments (e.g., early endosomes vs. plasma membrane), whereas in other tissues they localize to the same compartment (plasma membrane). We observed lysosomal co-localization between CATP-6::GFP and LMP-1::RFP in transgenic animals; however, this was an artifact of the tagged LMP-1 protein, since anti-LMP-1 antibody staining of native protein revealed that LMP-1 and CATP-6::GFP occupy different compartments. The nematode P5Bs are at least partially redundant, since we observed synthetic sterility in catp-5(0); catp-6(0) and catp-6(0) catp-7(0) double mutants. The double mutants exhibit defects in distal tip cell migration that resemble those of ina-1 (alpha integrin ortholog) and vab-3 (Pax6 ortholog) mutants, suggesting that the nematode P5Bs are required for ina-1and/or vab-3 function. This is potentially a conserved regulatory interaction, since mammalian ATP13A2, alpha integrin and Pax6 are all required for proper dopaminergic neuron function.

P5B ATPase存在于各种单细胞和多细胞真核生物的基因组中，表明它们具有悠久的起源，并且对细胞适应性很重要。ATP13A2（四种人类P5B ATP酶之一）的失活会导致帕金森氏病（库弗-拉克综合征）的早发。在跨膜区段M4的假定的底物相互作用口袋中存在不变的PPALP基序，这表明所有P5B ATPase可能具有相似的转运特异性。然而，运输基质的身份仍然未知。秀丽隐杆线虫的线虫具有三个旁系的P5B ATPase基因catp-5，catp-6和catp-7可能起源于Caenorhabditid进化枝附近的一个祖先基因。通过使用CRISPR / Cas9，我们系统地研究了C的每个P5B ATPase的表达模式，亚细胞定位和生物学功能。线虫。我们发现每个基因都有一个独特的表达模式，并且某些组织表达多个P5B。在表达模式重叠的某些组织中，不同的P5B靶向不同的亚细胞区室（例如，早期的内体与质膜），而在其他组织中，它们定位于相同的区室（质膜）。我们在转基因动物中观察到CATP-6 :: GFP和LMP-1 :: RFP之间的溶酶体共定位。但是，这是标记的LMP-1蛋白的伪影，因为天然蛋白的抗LMP-1抗体染色显示LMP-1和CATP-6 :: GFP占据不同的区室。线虫P5B至少部分冗余，因为我们在catp-5（0）中观察到合成的不育性。catp-6（0）和catp-6（0）catp-7（0）双突变体。双重突变体在远端末端细胞迁移中表现出缺陷，类似于ina-1（α整联蛋白直系同源物）和vab-3（Pax6直系同源物）突变体，这表明ina-1和/或vab-3功能需要线虫P5B。。这可能是保守的调节相互作用，因为哺乳动物的ATP13A2，α整联蛋白和Pax6都是正常的多巴胺能神经元功能所必需的。