Mutation in cpsf6/CFIm68 (Cleavage and Polyadenylation Specificity Factor Subunit 6) causes short 3'UTRs and disturbs gene expression in developing embryos, as revealed by an analysis of primordial germ cell migration using the medaka mutant naruto.
Mutation in cpsf6/CFIm68 (Cleavage and Polyadenylation Specificity Factor Subunit 6) causes short 3'UTRs and disturbs gene expression in developing embryos, as revealed by an analysis of primordial germ cell migration using the medaka mutant naruto.
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DOI：
10.1371/journal.pone.0172467
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Naruse K
中科院分区：
文献类型：
--
作者：
Sasado T;Kondoh H;Furutani-Seiki M;Naruse K
我们以前的研究分析青鳉突变体缺陷的原始生殖细胞（PGC）迁移确定cxcr 4 b和cxcr 7，这两个受体的趋化因子sdf 1/cxcl 12，作为关键的调节PGC迁移。在PGC迁移突变体中，鸣人（nar）是独特的，因为突变表型包括胚胎的总体形态异常，表明突变影响更广泛的过程。基因组测序结果表明，nar基因编码裂解和多聚腺苷酸化特异性因子亚基6（CPSF 6/CFIm 68）。CPSF 6是切割因子Im复合物（CFIm）的组分，其在前体mRNA 3 '-切割和聚腺苷酸化中起关键作用。突变体胚胎中sdf 1a/B和cxcr 7转录物的3 'RACE表明，与野生型胚胎相比，在更上游的位置处发生了更短的3' UTR和poly A添加，这表明CPSF 6具有防止过早的3 'UTR切割的功能。此外，在nar突变体中sdf 1 a/B的编码区序列的表达在体节中比野生型胚胎更向前延伸，解释了nar突变体中PGCs的异常延伸分布。缩短3 'UTR的预期结果是逃避由与远端3' UTR序列相互作用的microRNA介导的降解机制。sdf 1a编码序列的异常表达模式可能至少部分由这种机制解释。鉴于nar突变的多效性效应，使用nar突变体的进一步分析将揭示CPSF 6在多聚A位点选择中起重要调节作用的过程，以及3 'UTR参与体内各种基因的转录后基因调控。
Our previous studies analyzing medaka mutants defective in primordial germ cell (PGC) migration identified cxcr4b and cxcr7, which are both receptors of the chemokine sdf1/cxcl12, as key regulators of PGC migration. Among PGC migration mutants, naruto (nar) is unique in that the mutant phenotype includes gross morphological abnormalities of embryos, suggesting that the mutation affects a broader range of processes. A fine genetic linkage mapping and genome sequencing showed the nar gene encodes Cleavage and Polyadenylation Specificity Factor subunit 6 (CPSF6/CFIm68). CPSF6 is a component of the Cleavage Factor Im complex (CFIm) which plays a key role in pre-mRNA 3'-cleavage and polyadenylation. 3'RACE of sdf1a/b and cxcr7 transcripts in the mutant embryos indicated shorter 3’UTRs with poly A additions occurring at more upstream positions than wild-type embryos, suggesting CPSF6 functions to prevent premature 3’UTR cleavage. In addition, expression of the coding region sequences of sdf1a/b in nar mutants was more anteriorly extended in somites than wild-type embryos, accounting for the abnormally extended distribution of PGCs in nar mutants. An expected consequence of shortening 3'UTR is the escape from the degradation mechanism mediated by microRNAs interacting with distal 3’UTR sequence. The abnormal expression pattern of sdf1a coding sequence may be at least partially accounted for by this mechanism. Given the pleiotropic effects of nar mutation, further analysis using the nar mutant will reveal processes in which CPSF6 plays essential regulatory roles in poly A site selection and involvement of 3'UTRs in posttranscriptional gene regulation in various genes in vivo.