Braun Juliane, Meixner Arabella, Brachner Andreas, Foisner Roland
Max F. Perutz Laboratories (MFPL), Department of Medical Biochemistry, Medical University of Vienna, Vienna Biocenter (VBC), Vienna, Austria.
IMBA-Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, Austria.
PLoS One. 2016 Mar 24;11(3):e0152278. doi: 10.1371/journal.pone.0152278. eCollection 2016.
Ankyrin repeat and LEM-domain containing protein 1 (ANKLE1) is a GIY-YIG endonuclease with unknown functions, mainly expressed in mouse hematopoietic tissues. To test its potential role in hematopoiesis we generated Ankle1-deficient mice. Ankle1Δ/Δ mice are viable without any detectable phenotype in hematopoiesis. Neither hematopoietic progenitor cells, myeloid and lymphoid progenitors, nor B and T cell development in bone marrow, spleen and thymus, are affected in Ankle1Δ/Δ-mice. Similarly embryonic stress erythropoiesis in liver and adult erythropoiesis in bone marrow and spleen appear normal. To test whether ANKLE1, like the only other known GIY-YIG endonuclease in mammals, SLX1, may contribute to Holliday junction resolution during DNA repair, Ankle1-deficient cells were exposed to various DNA-damage inducing agents. However, lack of Ankle1 did not affect cell viability and, unlike depletion of Slx1, Ankle1-deficiency did not increase sister chromatid exchange in Bloom helicase-depleted cells. Altogether, we show that lack of Ankle1 does neither affect mouse hematopoiesis nor DNA damage repair in mouse embryonic fibroblasts, indicating a redundant or non-essential function of ANKLE1 in mouse.
锚蛋白重复序列和LEM结构域包含蛋白1（ANKLE1）是一种功能未知的GIY-YIG核酸内切酶，主要在小鼠造血组织中表达。为了测试其在造血过程中的潜在作用，我们构建了Ankle1基因敲除小鼠。Ankle1Δ/Δ小鼠能够存活，在造血方面没有任何可检测到的表型。Ankle1Δ/Δ小鼠的造血祖细胞、髓系和淋巴系祖细胞，以及骨髓、脾脏和胸腺中的B细胞和T细胞发育均未受到影响。同样，肝脏中的胚胎应激性红细胞生成以及骨髓和脾脏中的成年红细胞生成看起来也正常。为了测试ANKLE1是否像哺乳动物中另一种已知的GIY-YIG核酸内切酶SLX1一样，可能在DNA修复过程中促进霍利迪连接的解离，将Ankle1基因敲除的细胞暴露于各种DNA损伤诱导剂中。然而，缺乏Ankle1并不影响细胞活力，并且与Slx1缺失不同，Ankle1基因敲除不会增加布鲁姆解旋酶缺失细胞中的姐妹染色单体交换。总之，我们表明缺乏Ankle1既不影响小鼠造血，也不影响小鼠胚胎成纤维细胞中的DNA损伤修复，这表明ANKLE1在小鼠中具有冗余或非必需功能。
