Harris B, Dure L
Biochemistry. 1978 Aug 8;17(16):3250-6. doi: 10.1021/bi00609a012.
Evidence that RNA preexisting in the cotyledons of mature cotton seed (stored mRNA) is polyadenylated during the first day of germination is presented, based on three different experimental data sets. First, actinomycin D is found to inhibit 32PO4 incorporation into mRNA-poly(A) by 62%, into mRNA by 70%, but into poly(A) only 30%. Second, far more 32PO4 and [2-3H]adenosine are incorporated into the poly(A) portion of mRNA-poly(A) than into the mRNA portion as would be expected from their relative sizes and base composition. This underlabeling of the mRNA moiety is enhanced when cotyledons are germinated in actinomycin D. However, an expected distribution of the isotopes between the mRNA and poly(A) moieties is found in cotyledons labeled later in germination. Third, spectral measurements of the absolute amount of mRNA-poly(A) accumulated during the first day of germination in cotyledons germinated in actinomycin D are larger than would be expected from the 70% inhibition of mRNA labeling caused by the drug. The three sets of data suggest that over 50% of the total mass of mRNA polyadenylated during early germination exists in the mature seed. Its complexity, however, has not been measured. These data may explain the sensitivity of much of germination enzyme synthesis to inhibition by 3'dAdo during early germination and its insensitivity to actinomycin D during this period.
基于三个不同的实验数据集,有证据表明成熟棉籽子叶中预先存在的RNA(储存的mRNA)在萌发的第一天会发生多聚腺苷酸化。首先,发现放线菌素D可使32PO4掺入mRNA - poly(A)的量减少62%,掺入mRNA的量减少70%,但仅使掺入poly(A)的量减少30%。其次,与根据其相对大小和碱基组成所预期的情况相比,32PO4和[2 - 3H]腺苷掺入mRNA - poly(A)的poly(A)部分的量远多于掺入mRNA部分的量。当子叶在放线菌素D中萌发时,mRNA部分的这种低标记情况会增强。然而,在萌发后期标记的子叶中,发现了mRNA和poly(A)部分之间预期的同位素分布。第三,对在放线菌素D中萌发的子叶萌发第一天积累的mRNA - poly(A)绝对量的光谱测量结果,比根据该药物对mRNA标记的70%抑制作用所预期的结果要大。这三组数据表明,早期萌发过程中多聚腺苷酸化的mRNA总质量中超过50%存在于成熟种子中。然而,其复杂性尚未测量。这些数据可能解释了早期萌发过程中许多萌发酶合成对3'dAdo抑制的敏感性以及在此期间对放线菌素D的不敏感性。