Meister M, Braun A, Kappler C, Reichhart J M, Hoffmann J A
Institut de Biologie Moléculaire et Cellulaire, UPR 9022 du CNRS, Strasbourgh, France.
EMBO J. 1994 Dec 15;13(24):5958-66. doi: 10.1002/j.1460-2075.1994.tb06941.x.
Diptericins are antibacterial polypeptides which are strongly induced in the fat body and blood cells of dipteran insects in response to septic injury. The promoter of the single-copy, intronless diptericin gene of Drosophila contains several nucleotide sequences homologous to mammalian cis-regulatory motifs involved in the control of acute phase response genes. Extending our previous studies on the expression of the diptericin gene, we now report a quantitative analysis of the contribution of various putative regulatory elements to the bacterial inducibility of this gene, based on the generation of 60 transgenic fly lines carrying different elements fused to a reporter gene. Our data definitively identify two Kappa B-related motifs in the proximal promoter as the sites conferring inducibility and tissue-specific expression to the diptericin gene. These motifs alone, however, mediate only minimal levels of expression. Additional proximal regulatory elements are necessary to attain some 20% of the full response and we suspect a role for sequences homologous to mammalian IL6 response elements and interferon-gamma responsive sites in this up-regulation. The transgenic experiments also reveal the existence of a distal regulatory element located upstream of -0.6 kb which increases the level of expression by a factor of five.
双翅菌素是抗菌多肽,在双翅目昆虫受到败血症损伤时,会在其脂肪体和血细胞中强烈诱导产生。果蝇单拷贝、无内含子的双翅菌素基因的启动子包含几个核苷酸序列,这些序列与参与急性期反应基因调控的哺乳动物顺式调控基序同源。在之前对双翅菌素基因表达研究的基础上,我们现在报告一项定量分析,基于60个携带与报告基因融合的不同元件的转基因果蝇品系,分析各种假定调控元件对该基因细菌诱导性的贡献。我们的数据明确鉴定出近端启动子中的两个κB相关基序是赋予双翅菌素基因诱导性和组织特异性表达的位点。然而,仅这些基序只能介导最低水平的表达。还需要其他近端调控元件才能达到约20%的完全反应水平,我们怀疑与哺乳动物IL6反应元件和干扰素γ反应位点同源的序列在这种上调过程中发挥作用。转基因实验还揭示了位于-0.6 kb上游的一个远端调控元件的存在,它可使表达水平提高五倍。