Division of Translational Cancer Research, Department of Laboratory Medicine, Lund University, Lund, Sweden.
Lund Stem Cell Center, Department of Laboratory Medicine, Lund University, Lund, Sweden.
Sci Rep. 2018 Apr 23;8(1):6405. doi: 10.1038/s41598-018-24743-y.
KIT is a receptor tyrosine kinase (RTK) involved in several cellular processes such as regulation of proliferation, survival and differentiation of early hematopoietic cells, germ cells and melanocytes. Activation of KIT results in phosphorylation of tyrosine residues in the receptor, and recruitment of proteins that mediate downstream signaling and also modulate receptor signaling. Here we show that the SRC-like adaptor protein 2 (SLAP2) binds to wild-type KIT in a ligand-dependent manner and is furthermore found constitutively associated with the oncogenic mutant KIT-D816V. Peptide fishing analysis mapped pY568 and pY570 as potential SLAP2 association sites in KIT, which overlaps with the SRC binding sites in KIT. Expression of SLAP2 in cells expressing the transforming mutant KIT-D816V led to reduced cell viability and reduced colony formation. SLAP2 also partially blocked phosphorylation of several signal transduction molecules downstream of KIT such as AKT, ERK, p38 and STAT3. Finally, SLAP2 expression enhanced ubiquitination of KIT and its subsequent degradation. Taken together, our data demonstrate that SLAP2 negatively modulates KIT-D816V-mediated transformation by enhancing degradation of the receptor.
KIT 是一种受体酪氨酸激酶(RTK),参与多种细胞过程,如调节早期造血细胞、生殖细胞和黑素细胞的增殖、存活和分化。KIT 的激活导致受体中酪氨酸残基的磷酸化,并募集介导下游信号转导的蛋白,同时也调节受体信号转导。在这里,我们表明 SRC 样衔接蛋白 2(SLAP2)以配体依赖的方式与野生型 KIT 结合,并且还与致癌突变体 KIT-D816V 组成性相关。肽钓取分析将 pY568 和 pY570 映射为 KIT 中潜在的 SLAP2 结合位点,这与 KIT 中的 SRC 结合位点重叠。在表达转化突变体 KIT-D816V 的细胞中表达 SLAP2 导致细胞活力降低和集落形成减少。SLAP2 还部分阻断了 KIT 下游几种信号转导分子的磷酸化,如 AKT、ERK、p38 和 STAT3。最后,SLAP2 表达增强了 KIT 的泛素化及其随后的降解。总之,我们的数据表明,SLAP2 通过增强受体的降解来负调控 KIT-D816V 介导的转化。