Dreesen T D, Lezzi M, Case S T
Department of Biochemistry, University of Mississippi Medical Center, Jackson 39216-4505.
J Cell Biol. 1988 Jan;106(1):21-7. doi: 10.1083/jcb.106.1.21.
The expression of a Balbiani ring 1 gene that codes for a salivary gland-specific 180-kD secretory polypeptide (sp180) is regulated developmentally. Immunoblots of salivary gland protein incubated with an affinity-purified nonapeptide-reactive antibody demonstrated that the salivary gland content of sp180 increases as much as 10-fold between stages 8 and 10 of the fourth larval instar. Hybridization of RNA dot-blots with an oligonucleotide probe indicated that the observed increase in sp180 was preceded by a parallel 20-fold increase in the steady state level of its mRNA beginning between stages 7 and 8. In vitro nuclear transcription experiments demonstrated that there was a 10-fold acceleration in the rate of sp180 gene transcription between stages 6 and 10. The limited period of expression of the sp180 gene contrasted dramatically with the expression of Balbiani ring genes BR1, BR2 alpha, BR2 beta, and BR6, which code for the sp-I family of fibrous secretory polypeptides. The appearance of sp180 in secretion coincided with microscopically visible changes in the bundling of these fibrous polypeptides. At the same time, we noticed changes in the appearance and consistency of feeding tubes that larvae construct with this secretion. These results lead us to propose that sp180 may modify the structure or utilization of fibrous secretory polypeptides specifically for the assembly of pupation tubes necessary for larval/pupal ecdysis.
编码唾液腺特异性180-kD分泌多肽(sp180)的巴尔比亚尼环1基因的表达受到发育调控。用亲和纯化的九肽反应性抗体孵育唾液腺蛋白的免疫印迹表明,在第四龄幼虫的第8阶段和第10阶段之间,sp180的唾液腺含量增加了多达10倍。用寡核苷酸探针进行RNA点杂交表明,在第7阶段和第8阶段之间,其mRNA的稳态水平平行增加了20倍,随后才观察到sp180的增加。体外核转录实验表明,在第6阶段和第10阶段之间,sp180基因的转录速率加快了10倍。sp180基因的有限表达期与编码纤维状分泌多肽sp-I家族的巴尔比亚尼环基因BR1、BR2α、BR2β和BR6的表达形成了鲜明对比。sp180在分泌物中的出现与这些纤维状多肽的成束在显微镜下可见的变化同时发生。与此同时,我们注意到幼虫用这种分泌物构建的取食管的外观和稠度发生了变化。这些结果使我们提出,sp180可能专门修饰纤维状分泌多肽的结构或利用方式,以用于幼虫/蛹蜕皮所需的化蛹管的组装。