McCracken Allen, Locke John
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
PLoS One. 2014 Mar 10;9(3):e71695. doi: 10.1371/journal.pone.0071695. eCollection 2014.
Genes in multicellular organisms are expressed as part of a developmental program that is largely dependent on self-perpetuating higher-order chromatin states. The mechanism of establishing and maintaining these epigenetic events is well studied in Drosophila. The first known example of an epigenetic effect was that of (PEV) in Drosophila, which has been shown to be due to gene silencing via heterochromatin formation. We are investigating a process similar to Position Effect Variegation (PEV) using a mini-w transgene, called Pci, inserted in the upstream regulatory region of ci. The mini-white+ transgene in Pci is expressed throughout the adult eye; however, when other P or KP elements are present, a variegated eye phenotype results indicating random w+ silencing during development. This P element dependent silencing (PDS) can be modified by the haplo-suppressors/triplo-enhancers, Su(var)205 and Su(var)3-7, indicating that these heterochromatic modifiers also act dose dependently in PDS. Here we use a spontaneous derivative mutation of Pci called PciE1 (E1) that variegates like PDS in the absence of P elements, presumably due to an adjacent gypsy element insertion, to screen for second-site modifier mutations that enhance variable silencing of white+ in E1. We isolated 7 mutations in CG8878, an essential gene, that enhance the E1 variegated phenotype. CG8878, a previously uncharacterized gene, potentially encodes a serine/threonine kinase whose closest Drosophila paralogue, ballchen (nhk-1), phosphorylates histones. These mutant alleles enhance both PDS at E1 and Position Effect Variegation (PEV) at w(m4), indicating a previously unknown common silencing mechanism between the two.
多细胞生物中的基因作为发育程序的一部分被表达,该发育程序在很大程度上依赖于自我延续的高阶染色质状态。在果蝇中,建立和维持这些表观遗传事件的机制已得到充分研究。表观遗传效应的第一个已知例子是果蝇中的位置效应斑驳(PEV),已证明这是由于通过异染色质形成导致的基因沉默。我们正在使用插入到ci上游调控区域的一个称为Pci的mini-w转基因,研究一个类似于位置效应斑驳(PEV)的过程。Pci中的mini-white+转基因在整个成虫眼中都有表达;然而,当存在其他P或KP元件时,会产生斑驳的眼睛表型,表明在发育过程中w+随机沉默。这种P元件依赖性沉默(PDS)可以被单倍体抑制子/三倍体增强子Su(var)205和Su(var)3-7修饰,表明这些异染色质修饰因子在PDS中也呈剂量依赖性作用。在这里,我们使用Pci的一个自发衍生突变体PciE1(E1),它在没有P元件的情况下表现出类似于PDS的斑驳现象,推测是由于相邻的gypsy元件插入,来筛选增强E1中white+可变沉默的第二位点修饰突变。我们在一个必需基因CG8878中分离到了7个突变,这些突变增强了E1的斑驳表型。CG8878是一个先前未被表征的基因,可能编码一种丝氨酸/苏氨酸激酶,其在果蝇中最接近的旁系同源物ballchen(nhk-1)可磷酸化组蛋白。这些突变等位基因增强了E1处的PDS和w(m4)处的位置效应斑驳(PEV),表明两者之间存在一种先前未知的共同沉默机制。