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Quantitative analysis reveals genotype- and domain- specific differences between mRNA and protein expression of segmentation genes in Drosophila

机译:定量分析揭示了果蝇中分段基因的mRNA和蛋白质表达之间的基因型和结构域特异性差异

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In many biological systems gene expression at mRNA and protein levels is not identical. Rigorous comparison of such differences on a spatio-temporal scale is still not feasible by high-throughput transcriptomic and proteomic analyses of early embryo development. Here, we characterize differences between mRNA and protein expression of Drosophila segmentation genes at the level of individual gene expression domains. We obtained quantitative imaging data on expression of gap genes gt and hb and pair-rule gene eve for Drosophila wild type embryos, Kr null mutants and Kr+/Kr- heterozygotes. To compare mRNA and protein expression we use several criteria including difference in amplitude and positions of expression domains, pattern shape and positional variability. For a number of gene expression domains we show examples where protein expression does not repeat mRNA expression even after a temporal delay. We calculated time delays between eve pattern formation at the level of mRNA and protein for wild type embryos, Kr mutants and Kr+/Kr- heterozygotes. We detect that in wild type embryos, the amplitudes of eve stripes 3 and 7 do not differ significantly at the level of mRNA, however, stripe 3 is higher than stripe 7 at the protein level. We further show that hb mRNA and protein expression in both anterior and posterior domains significantly differs at specific time points. The formation of hb PS4 stripe at the mRNA level proceeds five times faster than at the level of protein. With regard to spatial expression, we show that the offset between posterior gt mRNA and protein domains is much larger in Kr mutants than in wild type embryos and heterozygotes. Finally, we analyze differences in positional variability of eve stripe 7 expression in Kr mutants and Kr+/Kr- heterozygotes at the level of mRNA and protein. These results enable further perspectives to uncover mechanisms underlying discrepancies between mRNA and protein expression in early embryo.
机译:在许多生物学系统中,mRNA和蛋白质水平的基因表达是不相同的。对时空规模的这种差异的严格比较仍然是通过早期胚胎发育的高通量转录组和蛋白质组学分析仍然不可行。在此,我们在单个基因表达结构域的水平下表征果蝇分段基因的mRNA和蛋白表达的差异。我们获得了用于果蝇野生型胚胎,KR NULL突变体和KR + / KR-杂合子的间隙基因GT和HB的表达和对规则基因EVE的定量成像数据。为了比较mRNA和蛋白质表达,我们使用若干标准,包括表达域,图案形状和位置变异性的幅度和位置的差异。对于许多基因表达结构域,我们表明即使在时间延迟之后,蛋白质表达不重复mRNA表达的实例。我们计算了野生型胚胎,KR突变体和KR + / Kr-杂合子的mRNA和蛋白水平的前夕模式形成的时间延迟。我们发现在野生型胚胎中,EVE条纹3和7的幅度在mRNA的水平下没有显着差异,然而,条纹3高于蛋白质水平的条纹7。我们进一步表明,在特定时间点,前部和后域的HB mRNA和蛋白质表达显着不同。在mRNA水平上形成Hb PS4条纹的速度比蛋白质水平快五倍。关于空间表达,我们表明,后GT mRNA和蛋白质结构域之间的偏移在KR突变体中大于野生型胚胎和杂合子。最后,我们分析在kr突变体和Kr + / kr-杂合中的eve条纹7表达的位置变异性的差异在mRNA和蛋白水平。这些结果使得能够进一步观察到揭示早期胚胎mRNA和蛋白质表达之间的差异差异的机制。

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