Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Heidelberg, Germany.
PLoS One. 2013 Oct 22;8(10):e77371. doi: 10.1371/journal.pone.0077371. eCollection 2013.
PUF proteins are a conserved family of RNA binding proteins found in all eukaryotes examined so far. This study focussed on PUF5, one of 11 PUF family members encoded in the Trypanosoma brucei genome. Native PUF5 is present at less than 50000 molecules per cell in both bloodstream and procyclic form trypanosomes. C-terminally myc-tagged PUF5 was mainly found in the cytoplasm and could be cross-linked to RNA. PUF5 knockdown by RNA interference had no effect on the growth of bloodstream forms. Procyclic forms lacking PUF5 grew normally, but expression of PUF5 bearing a 21 kDa tandem affinity purification tag inhibited growth. Knockdown of PUF5 did not have any effect on the ability of trypanosomes to differentiate from the mammalian to the insect form of the parasite.
PUF 蛋白是一个在迄今为止所有被研究过的真核生物中都存在的保守的 RNA 结合蛋白家族。本研究集中于 PUF5，这是在布氏锥虫基因组中编码的 11 个 PUF 家族成员之一。在血液体和前鞭毛体的寄生虫中，每个细胞中存在的天然 PUF5 分子少于 50000 个。C 端 myc 标记的 PUF5 主要存在于细胞质中，并可以与 RNA 交联。RNA 干扰引起的 PUF5 敲低对血液体形式的生长没有影响。缺乏 PUF5 的前鞭毛体正常生长，但携带 21 kDa 串联亲和纯化标签的 PUF5 表达抑制了生长。PUF5 的敲低对寄生虫从哺乳动物形式向昆虫形式分化的能力没有任何影响。
