期刊
VIRUSES-BASEL
卷 6, 期 4, 页码 1612-1636出版社
MDPI
DOI: 10.3390/v6041612
关键词
superresolution microscopy; mitochondria; MAM; MSIM; PALM; confocal microscopy; matrix; HCMV vMIA; GSTED; OMM
类别
资金
- NSF grant [MCB 1244509]
- core grant [1P30HD40677]
- Intramural Research Program of the National Institutes of Health, National Institute of Biomedical Imaging and Bioengineering
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1244509] Funding Source: National Science Foundation
The human cytomegalovirus (HCMV) viral mitochondria-localized inhibitor of apoptosis (vMIA) protein, traffics to mitochondria-associated membranes (MAM), where the endoplasmic reticulum (ER) contacts the outer mitochondrial membrane (OMM). vMIA association with the MAM has not been visualized by imaging. Here, we have visualized this by using a combination of confocal and superresolution imaging. Deconvolution of confocal microscopy images shows vMIA localizes away from mitochondrial matrix at the Mitochondria-ER interface. By gated stimulated emission depletion (GSTED) imaging, we show that along this interface vMIA is distributed in clusters. Through multicolor, multifocal structured illumination microscopy (MSIM), we find vMIA clusters localize away from MitoTracker Red, indicating its OMM localization. GSTED and MSIM imaging show vMIA exists in clusters of ~100-150 nm, which is consistent with the cluster size determined by Photoactivated Localization Microscopy (PALM). With these diverse superresolution approaches, we have imaged the clustered distribution of vMIA at the OMM adjacent to the ER. Our findings directly compare the relative advantages of each of these superresolution imaging modalities for imaging components of the MAM and sub-mitochondrial compartments. These studies establish the ability of superresolution imaging to provide valuable insight into viral protein location, particularly in the sub-mitochondrial compartments, and into their clustered organization.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据