期刊
TRAFFIC
卷 9, 期 6, 页码 995-1018出版社
WILEY
DOI: 10.1111/j.1600-0854.2008.00732.x
关键词
early endosome; EHD1; EHD4; Rab5
类别
资金
- NATIONAL CENTER FOR RESEARCH RESOURCES [P20RR018759] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM087455, R01GM074876] Funding Source: NIH RePORTER
- NCRR NIH HHS [P20 RR018759, 5P20 RR018759-04] Funding Source: Medline
- NIGMS NIH HHS [1R01GM074876, R01 GM074876-04, R01 GM087455, R01 GM087455-02, R01 GM074876] Funding Source: Medline
All four of the C-terminal Eps15 homology domain (EHD) proteins have been implicated in the regulation of endocytic trafficking. However, the high level of amino acid sequence identity among these proteins has made it challenging to elucidate the precise function of individual EHD proteins. We demonstrate here with specific peptide antibodies that endogenous EHD4 localizes to Rab5-, early embryonic antigen 1 (EEA1)- and Arf6-containing endosomes and colocalizes with internalized transferrin in the cell periphery. Knock-down of EHD4 expression by both small interfering RNA and short hairpin RNA leads to the generation of enlarged early endosomal structures that contain Rab5 and EEA1 as well as internalized transferrin or major histocompatibility complex class I molecules. In addition, cargo destined for degradation, such as internalized low-density lipoprotein, also accumulates in the enlarged early endosomes in EHD4-depleted cells. Moreover, we have demonstrated that these enlarged early endosomes are enriched in levels of the activated GTP-bound Rab5. Finally, we show that endogenous EHD4 and EHD1 interact in cells, suggesting coordinated involvement in the regulation of receptor transport along the early endosome to endocytic recycling compartment axis. The results presented herein provide evidence that EHD4 is involved in the control of trafficking at the early endosome and regulates exit of cargo toward both the recycling compartment and the late endocytic pathway.
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