Ritter L M, Bryans M, Abdo O, Sharma V, Wilkie N M
Department of Molecular Genetics, Ohio State University, Columbus, USA.
Mol Cell Biol. 1995 Jun;15(6):3110-8. doi: 10.1128/MCB.15.6.3110.
Murine macrophage inflammatory protein 1 alpha (MIP-1 alpha) and its human equivalent (GOS19, LD78, or AT464) are members of the -C-C family of low-molecular-weight chemokines. Secreted from activated T cells and macrophages, bone marrow-derived MIP-1 alpha/GOS19 inhibits primitive hematopoietic stem cells and appears to be involved in the homeostatic control of stem cell proliferation. It also induces chemotaxis and inflammatory responses in mature cell types. Therefore, it is important to understand the mechanisms which control the expression of MIP-1 alpha/GOS19. Previous work has shown that in Jurkat T cells, a set of widely expressed transcription factors (the ICK-1 family) affect the GOS19 promoter. One member, ICK-1A, behaves as a strong negative regulator. In this communication, we provide evidence that the pathway of induction in the macrophage cell line U937 is different from that in Jurkat cells. Furthermore, we show that the ICK-1 binding site does not confer negative regulation in U937 cells. We provide evidence for an additional binding site, the MIP-1 alpha nuclear protein (MNP) site, which overlaps the ICK-1 site. Interaction of nuclear extracts from various cell lines and tissue with the MNP site leads to the formation of fast-migrating protein-DNA complexes with similar but distinct electrophoretic mobilities. A mutation of the MNP site which does not abrogate ICK-1 binding inactivates the GOS19.1 promoter in U937 cells and reduces its activity by fourfold in Jurkat cells. We propose that the MNP protein(s) binding at the MNP site constitutes a novel transcription factor(s) expressed in hematopoietic cells.
小鼠巨噬细胞炎性蛋白1α(MIP-1α)及其人类同源物(GOS19、LD78或AT464)是低分子量趋化因子-C-C家族的成员。骨髓来源的MIP-1α/GOS19由活化的T细胞和巨噬细胞分泌,可抑制原始造血干细胞,似乎参与干细胞增殖的稳态控制。它还能诱导成熟细胞类型的趋化性和炎症反应。因此,了解控制MIP-1α/GOS19表达的机制很重要。先前的研究表明,在Jurkat T细胞中,一组广泛表达的转录因子(ICK-1家族)影响GOS19启动子。其中一个成员ICK-1A表现为强负调节因子。在本通讯中,我们提供证据表明巨噬细胞系U937中的诱导途径与Jurkat细胞中的不同。此外,我们表明ICK-1结合位点在U937细胞中不具有负调节作用。我们提供了一个额外结合位点的证据,即MIP-1α核蛋白(MNP)位点,它与ICK-1位点重叠。各种细胞系和组织的核提取物与MNP位点的相互作用导致形成具有相似但不同电泳迁移率的快速迁移的蛋白质-DNA复合物。一个不消除ICK-1结合的MNP位点突变使U937细胞中的GOS19.1启动子失活,并使Jurkat细胞中的活性降低四倍。我们提出,结合在MNP位点的MNP蛋白构成了一种在造血细胞中表达的新型转录因子。