聚（ADP - 核糖）调节MARCKS蛋白的特性。

Poly(ADP-ribose) modulates the properties of MARCKS proteins.

作者信息

Schmitz A A, Pleschke J M, Kleczkowska H E, Althaus F R, Vergères G

机构信息

Department of Biophysical Chemistry, Biozentrum, University of Basel, Switzerland.

出版信息

Biochemistry. 1998 Jun 30;37(26):9520-7. doi: 10.1021/bi973063b.

DOI:10.1021/bi973063b

PMID:9649335

Abstract

In mammalian cells, the formation of DNA strand breaks is accompanied by synthesis of poly(ADP-ribose). This nucleic acid-like homopolymer may modulate protein functions by covalent and/or noncovalent interactions. Here we show that poly(ADP-ribose) binds strongly to the proteins of the myristoylated alanine-rich C kinase substrate (MARCKS) family, MARCKS and MARCKS-related protein (also MacMARCKS or F52). MARCKS proteins are myristoylated proteins associated with membranes and the actin cytoskeleton. As targets for both protein kinase C (PKC) and calmodulin (CaM), MARCKS proteins are thought to mediate cross-talk between these two signal transduction pathways. Dot blot assays show that poly(ADP-ribose) binds to MARCKS proteins at the highly basic effector domain. Complex formation between MARCKS-related protein and CaM as well as phosphorylation of MARCKS-related protein by the catalytic subunit of PKC are strongly inhibited by equimolar amounts of poly(ADP-ribose), suggesting a high affinity of poly(ADP-ribose) for MARCKS-related protein. Binding of MARCKS-related protein to membranes is also inhibited by poly(ADP-ribose). Finally, poly(ADP-ribose) efficiently reverses the actin-filament bundling activity of a peptide corresponding to the effector domain and inhibits the formation of actin filaments in vitro. Our results suggest that MARCKS proteins and actin could be targets of the poly(ADP-ribose) DNA damage signal pathway.

摘要

在哺乳动物细胞中，DNA链断裂的形成伴随着聚（ADP - 核糖）的合成。这种类似核酸的同聚物可通过共价和/或非共价相互作用调节蛋白质功能。在此我们表明，聚（ADP - 核糖）与富含肉豆蔻酰化丙氨酸的C激酶底物（MARCKS）家族的蛋白质、MARCKS和MARCKS相关蛋白（也称为MacMARCKS或F52）紧密结合。MARCKS蛋白是与膜和肌动蛋白细胞骨架相关的肉豆蔻酰化蛋白。作为蛋白激酶C（PKC）和钙调蛋白（CaM）的靶点，MARCKS蛋白被认为介导这两条信号转导途径之间的相互作用。斑点印迹分析表明，聚（ADP - 核糖）在高度碱性的效应器结构域与MARCKS蛋白结合。等摩尔量的聚（ADP - 核糖）强烈抑制MARCKS相关蛋白与CaM之间的复合物形成以及PKC催化亚基对MARCKS相关蛋白的磷酸化，这表明聚（ADP - 核糖）对MARCKS相关蛋白具有高亲和力。聚（ADP - 核糖）也抑制MARCKS相关蛋白与膜的结合。最后，聚（ADP - 核糖）有效地逆转了与效应器结构域对应的肽段的肌动蛋白丝束集活性，并在体外抑制肌动蛋白丝的形成。我们的结果表明，MARCKS蛋白和肌动蛋白可能是聚（ADP - 核糖）DNA损伤信号通路的靶点。