Correlated cryogenic fluorescence microscopy and electron cryo-tomography shows that exogenous TRIM5α can form hexagonal lattices or autophagy aggregates in vivo

被引:18
作者
Carter, Stephen D. [1 ]
Mamede, Joao, I [2 ,3 ]
Hope, Thomas J. [2 ]
Jensen, Grant J. [1 ,4 ]
机构
[1] CALTECH, Div Biol & Biol Engn, Pasadena, CA 91125 USA
[2] Northwestern Univ, Dept Cell & Dev Biol, Chicago, IL 60611 USA
[3] Rush Univ, Med Ctr, Dept Microbial Pathogens & Immun, Chicago, IL 60612 USA
[4] CALTECH, HHMI, Pasadena, CA 91125 USA
关键词
cryo-CLEM/ECT; TRIM5; alpha-body; hexagonal nets; autophagy; endoplasmic reticulum; PROTEINS REGULATE AUTOPHAGY; UBIQUITINATED PROTEINS; TRANSIENT AGGREGATION; RESTRICTION; HIV-1; PROTEASOME; VISUALIZATION; P62/SQSTM1; COMPARTMENTS; RECOGNITION;
D O I
10.1073/pnas.1920323117
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Members of the tripartite motif (TRIM) protein family have been shown to assemble into structures in both the nucleus and cytoplasm. One TRIM protein family member, TRIM5 alpha, has been shown to form cytoplasmic bodies involved in restricting retroviruses such as HIV-1. Here we applied cryogenic correlated light and electron microscopy, combined with electron cryo-tomography, to intact mammalian cells expressing YFP-rhTRIM5 alpha and found the presence of hexagonal nets whose arm lengths were similar to those of the hexagonal nets formed by purified TRIM5 alpha in vitro. We also observed YFP-rhTRIM5 alpha within a diversity of structures with characteristics expected for organelles involved in different stages of macroautophagy, including disorganized protein aggregations (sequestosomes), sequestosomes flanked by flat double-membraned vesicles (sequestosome:phagophore complexes), sequestosomes within double-membraned vesicles (autophagosomes), and sequestosomes within multivesicular autophagic vacuoles (amphisomes or autolysosomes). Vaults were also seen in these structures, consistent with their role in autophagy. Our data 1) support recent reports that TRIM5 alpha can form both well-organized signaling complexes and nonsignaling aggregates, 2) offer images of the macroautophagy pathway in a near-native state, and 3) reveal that vaults arrive early in macroautophagy.
引用
收藏
页码:29702 / 29711
页数:10
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