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FASEB J. 2014 Jan;28(1):106-16. doi: 10.1096/fj.13-238113. Epub 2013 Oct 4.
A 27-aa peptide (P27) was previously shown to decrease the accumulation of human immunodeficiency virus type 1 (HIV-1) in the supernatant of chronically infected cells; however, the mechanism was not understood. Here, we show that P27 prevents virus accumulation by inducing macropinocytosis (MPC). Treatment of HIV-1- and human T-cell lymphotropic virus type 1 (HTLV-1)-infected cells with 2-10 μM P27 caused cell membrane ruffling and uptake of virus and polymerized forms of the peptide into large vacuoles. As demonstrated by electron microscopy, activation of MPC did not require virus or cells infected with virus, as P27 initiated its own uptake in the absence of virus. Inhibitors of MPC, Cytochalasin D and amiloride, decreased P27-mediated uptake of soluble dextran and inhibited P27-induced virus uptake by >60%, which provides further evidence that P27 induces MPC. In CD4(+) HeLa cells, HIV-1 infection was enhanced by P27 up to 4-fold, and P27 increased infection at concentrations as low as 20 nM. The 5-aa C-terminal domain of P27 was necessary for virus uptake and may be responsible for the polymerization of P27 into fibrils. These forms of P27 may play a key role in triggering MPC, making this peptide a useful tool for studying virus uptake and infection, as well as MPC of other macromolecules.
先前的研究表明,一种 27 个氨基酸组成的肽(P27)可减少慢性感染细胞上清液中人类免疫缺陷病毒 1 型(HIV-1)的积累,但具体机制尚未阐明。本文研究发现,P27 通过诱导巨胞饮作用(MPC)来防止病毒积累。用 2-10 μM P27 处理 HIV-1 和人嗜 T 淋巴细胞病毒 1 型(HTLV-1)感染的细胞,可导致细胞膜起皱,并将病毒和肽的聚合形式摄入大空泡。电子显微镜显示,MPC 的激活不需要病毒或感染病毒的细胞,因为 P27 在没有病毒的情况下可自行内化。MPC 的抑制剂细胞松弛素 D 和阿米洛利,降低了 P27 介导的可溶性葡聚糖的摄取,并抑制了 P27 诱导的病毒摄取超过 60%,这进一步证明 P27 诱导了 MPC。在 CD4(+) HeLa 细胞中,P27 可将 HIV-1 感染提高 4 倍,且 P27 在低至 20 nM 的浓度下即可增加感染。P27 的 5 个氨基酸 C 末端结构域是病毒摄取所必需的,可能负责 P27 聚合形成原纤维。这些形式的 P27 可能在触发 MPC 中起关键作用,使该肽成为研究病毒摄取和感染以及其他大分子 MPC 的有用工具。