Special Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi, India.
Special Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi, India.
J Biol Chem. 2022 Dec;298(12):102646. doi: 10.1016/j.jbc.2022.102646. Epub 2022 Oct 26.
Iron (Fe) sequestration is one of the most important strategies of the host to control the growth and survival of invading pathogens. Ferritin (Ft) plays a pivotal role in the sequestration mechanism of mammalian hosts by storing Fe. Recent evidence suggests that poly(rC)-binding proteins (PCBPs) act as chaperones for loading Fe into Ft. Incidentally, modulation of host PCBPs in respect to storing Fe in Ft during any infection remains unexplored. Among PCBPs, PCBP1 and PCBP2 are present in every cell type and involved in interacting with Ft for Fe loading. Leishmania donovani (LD) resides within macrophages during the mammalian stage of infection, causing life-threatening visceral leishmaniasis. Here, we reveal the ability of LD to cleave PCBP1 and PCBP2 in host monocytes/macrophages. LD cleaves PCBP1-FLAG into two fragments and PCBP2-FLAG into multiple fragments, thus affecting their interactions with Ft and resulting in decreased Fe loading into Ft. LD-derived culture supernatant or exosome-enriched fractions are also able to cleave PCBPs, suggesting involvement of a secreted protease of the parasite. Using an immune-depletion experiment and transgenic mutants, we confirmed the involvement of zinc-metalloprotease GP63 in cleaving PCBPs. We further revealed that by cleaving host PCBPs, Leishmania could inhibit Fe loading into Ft to accumulate available Fe for higher intracellular growth. This is the first report of a novel strategy adopted by a mammalian pathogen to interfere with Fe sequestration into Ft by cleaving chaperones for its survival advantage within the host.
铁(Fe)螯合是宿主控制入侵病原体生长和存活的最重要策略之一。铁蛋白(Ft)通过储存 Fe 在哺乳动物宿主的螯合机制中起着关键作用。最近的证据表明,多聚(C)结合蛋白(PCBP)作为将 Fe 加载到 Ft 中的伴侣蛋白发挥作用。顺便说一句,在任何感染期间,宿主 PCBP 的调节在将 Fe 储存在 Ft 中方面仍未得到探索。在 PCBP 中,PCBP1 和 PCBP2 存在于每种细胞类型中,并参与与 Ft 相互作用以加载 Fe。利什曼原虫(LD)在感染的哺乳动物阶段存在于巨噬细胞内,导致危及生命的内脏利什曼病。在这里,我们揭示了 LD 能够在宿主单核细胞/巨噬细胞中切割 PCBP1 和 PCBP2 的能力。LD 将 PCBP1-FLAG 切割成两个片段,将 PCBP2-FLAG 切割成多个片段,从而影响它们与 Ft 的相互作用,并导致 Fe 加载到 Ft 减少。LD 衍生的培养上清液或富含外泌体的级分也能够切割 PCBP,表明寄生虫的一种分泌蛋白酶参与其中。通过免疫耗竭实验和转基因突变体,我们证实锌金属蛋白酶 GP63 参与了 PCBP 的切割。我们进一步揭示,通过切割宿主 PCBP,利什曼原虫可以抑制 Fe 加载到 Ft 中,从而积累可用的 Fe 以促进细胞内生长。这是哺乳动物病原体采用的一种新策略的首次报道,该策略通过切割伴侣蛋白来干扰 Fe 螯合到 Ft 中,从而在宿主内获得生存优势。