Strand selective generation of endo-siRNAs from the Na/phosphate transporter gene Slc34a1 in murine tissues.
Strand selective generation of endo-siRNAs from the Na/phosphate transporter gene Slc34a1 in murine tissues.
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DOI：
10.1093/nar/gkp088
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发表时间：
2009-04
影响因子：
14.9
通讯作者：
Werner A
中科院分区：
文献类型：
--
作者：
Carlile M;Swan D;Jackson K;Preston-Fayers K;Ballester B;Flicek P;Werner A
天然反义转录物（NATs）是基因表达的重要调控因子。最近，反义转录和内切sirna形成之间的联系已经出现。我们从内切sirna加工的角度研究了Na/phosphate共转运体基因（Slc34a1）的双向转录。小鼠Slc34a1产生一个反义转录物，代表位于Slc34a1下游的Pfn3基因的另一种剪接产物。反义转录本在睾丸和肾脏中显著存在。体外合成的正义/反义转录物在非洲爪蟾卵母细胞中的共表达表明重叠转录物在细胞核中加工成内切sirna。截断实验表明，至少29个碱基对的重叠需要诱导加工。我们通过northern blotting检测了小鼠组织中共同表达Slc34a1正义/反义转录物的内切sirna。在小鼠肾脏和睾丸中，内切sirna的取向具有组织特异性。在肾脏中，Na/磷酸盐共转运体履行其生理功能，检测到与NAT互补的内腔sirna，在睾丸中发现了两个方向。考虑到NATs的广泛表达和内切sirna的基因沉默潜力，我们假设反义转录和单等位基因表达之间存在全基因组联系。随机印迹与反义转录之间确实存在显著的相关性。我们的研究结果表明，NATs在基因调控中具有一种新的、更普遍的作用。
Natural antisense transcripts (NATs) are important regulators of gene expression. Recently, a link between antisense transcription and the formation of endo-siRNAs has emerged. We investigated the bi-directionally transcribed Na/phosphate cotransporter gene (Slc34a1) under the aspect of endo-siRNA processing. Mouse Slc34a1 produces an antisense transcript that represents an alternative splice product of the Pfn3 gene located downstream of Slc34a1. The antisense transcript is prominently found in testis and in kidney. Co-expression of in vitro synthesized sense/antisense transcripts in Xenopus oocytes indicated processing of the overlapping transcripts into endo-siRNAs in the nucleus. Truncation experiments revealed that an overlap of at least 29 base-pairs is required to induce processing. We detected endo-siRNAs in mouse tissues that co express Slc34a1 sense/antisense transcripts by northern blotting. The orientation of endo-siRNAs was tissue specific in mouse kidney and testis. In kidney where the Na/phosphate cotransporter fulfils its physiological function endo-siRNAs complementary to the NAT were detected, in testis both orientations were found. Considering the wide spread expression of NATs and the gene silencing potential of endo-siRNAs we hypothesized a genome-wide link between antisense transcription and monoallelic expression. Significant correlation between random imprinting and antisense transcription could indeed be established. Our findings suggest a novel, more general role for NATs in gene regulation.
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影响因子：
11.4
作者：
Krueger, Christel;Morison, Ian M.
通讯作者：
Morison, Ian M.
DOI：
10.1073/pnas.0805781105
发表时间：
2008-09-16
影响因子：
11.1
作者：
Rudnick, Stephen I.;Swaminathan, Jyothishmathi;Gewirtz, Alan M.
通讯作者：
Gewirtz, Alan M.
影响因子：
56.9
作者：
Morris, KV;Chan, SWL;Looney, DJ
通讯作者：
Looney, DJ
影响因子：
5.5
作者：
Nalbant, P;Boehmer, C;Werner, A
通讯作者：
Werner, A
影响因子：
7.7
作者：
Lapidot, Michal;Pilpel, Yitzhak
通讯作者：
Pilpel, Yitzhak