Bellini M, Lacroix J C, Gall J G
Laboratoire de Génétique du Développement, Université Pierre et Marie Curie, Paris, France.
EMBO J. 1993 Jan;12(1):107-14. doi: 10.1002/j.1460-2075.1993.tb05636.x.
We used mAb A33/22, which recognizes a nuclear protein on the loops of amphibian lampbrush chromosomes, to select cDNA clone PwA33 from an expression library of the newt Pleurodeles waltl. A myc-tagged transcript of clone PwA33 was injected into Pleurodeles oocytes. The translation product localized in the germinal vesicle (GV) and was distributed on the lampbrush loops in a pattern identical to that of the endogenous protein. PwA33 encodes a 71 kDa protein with three distinct domains: a region rich in Cys/His residues that may form zinc fingers, a coiled-coil domain with potential for dimerization and a third 'rfp-like' domain that is shared by several other nuclear proteins. The putative zinc fingers and the coiled-coil domain resemble features in known nucleic acid-binding regulatory proteins. These structures, coupled with a distinctive pattern of expression in embryonic tissues, suggest that A33 may function as a regulatory protein during early development. It is unlikely that the large store of A33 in the GV is bound to DNA. Instead, its association with the nascent transcripts on the lampbrush chromosome loops suggests a role in pre-mRNA synthesis or processing.
我们使用识别两栖动物灯刷染色体环上一种核蛋白的单克隆抗体A33/22,从蝾螈(Pleurodeles waltl)的表达文库中筛选出cDNA克隆PwA33。将带有myc标签的克隆PwA33转录本注射到蝾螈卵母细胞中。翻译产物定位于生发泡(GV)中,并以与内源性蛋白相同的模式分布在灯刷环上。PwA33编码一种71 kDa的蛋白质,具有三个不同的结构域:一个富含半胱氨酸/组氨酸残基的区域,可能形成锌指结构;一个具有二聚化潜力的卷曲螺旋结构域;以及第三个“rfp样”结构域,该结构域为其他几种核蛋白所共有。推测的锌指结构和卷曲螺旋结构域类似于已知核酸结合调节蛋白中的特征。这些结构,再加上在胚胎组织中独特的表达模式,表明A33可能在早期发育过程中作为一种调节蛋白发挥作用。生发泡中大量储存的A33不太可能与DNA结合。相反,它与灯刷染色体环上新生转录本的结合表明其在mRNA前体合成或加工中发挥作用。