Journal
MOLECULAR BIOLOGY OF THE CELL
Volume 27, Issue 1, Pages 108-126Publisher
AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E15-07-0514
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Funding
- National Institutes of Health [R01GM087455, R01GM074876]
- Nebraska Research Initiative
- Fred and Pamela Buffett Cancer Center Support Grant [P30CA036727]
- National Institute of General Medical Sciences of the National Institutes of Health [P30GM106397]
- National Institutes of Health
- NATIONAL CANCER INSTITUTE [P30CA036727] Funding Source: NIH RePORTER
- NATIONAL CENTER FOR RESEARCH RESOURCES [P20RR018759] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [P30GM106397, R01GM087455, R01GM074876] Funding Source: NIH RePORTER
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The endocytic recycling compartment (ERC) is a series of perinuclear tubular and vesicular membranes that regulates recycling to the plasma membrane. Despite evidence that cargo is sorted at the early/sorting endosome (SE), whether cargo mixes downstream at the ERC or remains segregated is an unanswered question. Here we use three-dimensional (3D) structured illumination microscopy and dual-channel and 3D direct stochastic optical reconstruction microscopy (dSTORM) to obtain new information about ERC morphology and cargo segregation. We show that cargo internalized either via clathrin-mediated endocytosis (CME) or independently of clathrin (CIE) remains segregated in the ERC, likely on distinct carriers. This suggests that no further sorting occurs upon cargo exit from SE. Moreover, 3D dSTORM data support a model in which some but not all ERC vesicles are tethered by contiguous membrane bridges. Furthermore, tubular recycling endosomes preferentially traffic CIE cargo and may originate from SE membranes. These findings support a significantly altered model for endocytic recycling in mammalian cells in which sorting occurs in peripheral endosomes and segregation is maintained at the ERC.
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