Conlon R A, Tufaro F, Brandhorst B P
Department of Biology, McGill University, Montreal, P.Q., Canada.
Genes Dev. 1987 Jun;1(4):337-46. doi: 10.1101/gad.1.4.337.
The synthesis of many paternal species-specific proteins is reduced in all stages of sea urchin interspecies hybrid embryos, due to the reduced amounts of some paternal mRNA species in hybrid embryos compared with the embryos of the paternal species (Tufaro and Brandhorst 1982). Possible explanations for this restriction were tested. Cloned cDNAs were selected that were specific for paternal RNA sequences having reduced amounts (to 2-20% of normal) in hybrid embryos derived from a cross of Stronglyocentrotus purpuratus eggs with Lytechinus pictus sperm. Several of these RNA species are barely detectable in the eggs, but they accumulate extensively (5- to 40-fold) during L. pictus embryogenesis. Thus, the restricted expression of these paternal genes in hybrid embryos is not the result of the persistence of stable maternal mRNA species stored in eggs and not replaced by zygotic transcription. The accumulation of some of these L. pictus transcripts is also reduced in the reciprocal cross (L. pictus eggs X S. purpuratus sperm); therefore, the full expression of these L. pictus genes in hybrid embryos is not dependent on species-specific maternal factors stored in the egg. The transcriptional activity of one such gene was estimated using a run-on assay in isolated nuclei; it is as actively transcribed in hybrid as it is in homospecific embryos, but in hybrid embryos the cytoplasmic transcript accumulates to only 2-15% of the normal level. Sequence analysis indicates that this gene encodes a metallothionein. Mechanisms are discussed that might account for the post-transcriptional restriction of expression of some genes in hybrid embryos.
在海胆种间杂交胚胎的所有发育阶段,许多父本物种特异性蛋白质的合成都会减少,这是因为与父本物种的胚胎相比,杂交胚胎中某些父本mRNA种类的含量有所降低(图法罗和布兰德霍斯特,1982年)。针对这种限制的可能解释进行了测试。选择了一些克隆的cDNA,这些cDNA对来自紫海胆卵子与花斑海胆精子杂交产生的杂交胚胎中含量降低(降至正常水平的2%-20%)的父本RNA序列具有特异性。这些RNA种类中有几种在卵子中几乎检测不到,但它们在花斑海胆胚胎发育过程中大量积累(增加5至40倍)。因此,这些父本基因在杂交胚胎中的表达受限并非是由于卵子中储存的稳定母本mRNA种类持续存在且未被合子转录所取代。在反交(花斑海胆卵子×紫海胆精子)中,这些花斑海胆转录本中的一些积累也会减少;因此,这些花斑海胆基因在杂交胚胎中的完全表达并不依赖于卵子中储存的物种特异性母本因子。使用分离细胞核中的连续转录分析估计了其中一个这样的基因的转录活性;它在杂交胚胎中的转录活性与在同种胚胎中一样高,但在杂交胚胎中,细胞质转录本仅积累到正常水平的2%-15%。序列分析表明该基因编码一种金属硫蛋白。文中讨论了可能解释杂交胚胎中某些基因转录后表达受限的机制。