Depletion of the Trypanosome Pumilio Domain Protein PUF2 or of Some Other Essential Proteins Causes Transcriptome Changes Related to Coding Region Length
Depletion of the Trypanosome Pumilio Domain Protein PUF2 or of Some Other Essential Proteins Causes Transcriptome Changes Related to Coding Region Length
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DOI：
10.1128/ec.00018-14
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发表时间：
2014-05-01
期刊：
影响因子：
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通讯作者：
Clayton, Christine
中科院分区：
文献类型：
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作者：
Jha, Bhaskar Anand;Fadda, Abeer;Clayton, Christine
已知的Pumilio结构域RNA结合蛋白主要是转录后抑制基因表达，降低mRNA的翻译和稳定性。布氏锥虫有11种PUF蛋白。我们在这里展示了PUF2在细胞质中，每个细胞的分子数量与mRNAs大致相同。虽然PUF2在体内表现出低水平的RNA结合，但它与多聚体无关。PUF2也降低了报告基因的表达水平，这与抑制作用一致。PUF2的缺失抑制了血流状锥虫的生长，导致开放阅读框较长的mRNAs选择性丢失，开放阅读框较短的mRNAs增加。对已发表的RNAseq数据的重新检查发现，在缺乏一些其他蛋白质的细胞中也有同样的趋势。我们推测，这些长度效应可能是由于抑制了转录的延伸阶段，或者是由于翻译状态或多聚体构象对mRNA衰退的影响所致。
Pumilio domain RNA-binding proteins are known mainly as posttranscriptional repressors of gene expression that reduce mRNA translation and stability. Trypanosoma brucei has 11 PUF proteins. We show here that PUF2 is in the cytosol, with roughly the same number of molecules per cell as there are mRNAs. Although PUF2 exhibits a low level of in vivo RNA binding, it is not associated with polysomes. PUF2 also decreased reporter mRNA levels in a tethering assay, consistent with a repressive role. Depletion of PUF2 inhibited growth of bloodstream-form trypanosomes, causing selective loss of mRNAs with long open reading frames and increases in mRNAs with shorter open reading frames. Reexamination of published RNASeq data revealed the same trend in cells depleted of some other proteins. We speculate that these length effects could be caused by inhibition of the elongation phase of transcription or by an influence of translation status or polysomal conformation on mRNA decay.