TY - JOUR
T1 - HID-1 controls formation of large dense core vesicles by influencing cargo sorting and trans-Golgi network acidification
AU - Hummer, Blake H.
AU - De Leeuw, Noah F.
AU - Burns, Christian
AU - Chen, Lan
AU - Joens, Matthew S.
AU - Hosford, Bethany
AU - Fitzpatrick, James A.J.
AU - Asensio, Cedric S.
N1 - Funding Information:
We thank Emmanuel Boucrot and Dan Sirkis for critical reading of the manuscript, Dinah Loerke for help with image analysis, the An-gleson lab for access to their microscope, members of the Asensio lab for thoughtful discussions, Josh Loomis and Shirley Sobus for help with cell sorting and analysis at the National Jewish Health Center flow cytometry core facility (Denver), Xiao-Song Xie for the a2 antibody, and the National Institutes of Health (GM116096), the American Heart Association (#16BGIA27780049), and the American Diabetes Association (1-17-JDF-064) for support to C.S.A. M.S.J. and J.A.J.F. gratefully acknowledge support from the Washington University Center for Cellular Imaging, which is supported by the Washington University School of Medicine, the Children’s Discovery Institute of Washington University and St. Louis Children’s Hospital (CDI-CORE-2015-505), and the Foundation for Barnes Jewish Hospital.
Publisher Copyright:
© 2017 Hummer et al.
PY - 2017/12
Y1 - 2017/12
N2 - Large dense core vesicles (LDCVs) mediate the regulated release of neuropeptides and peptide hormones. They form at the trans-Golgi network (TGN), where their soluble content aggregates to form a dense core, but the mechanisms controlling biogenesis are still not completely understood. Recent studies have implicated the peripheral membrane protein HID-1 in neuropeptide sorting and insulin secretion. Using CRISPR/Cas9, we generated HID-1 KO rat neuroendocrine cells, and we show that the absence of HID-1 results in specific defects in peptide hormone and monoamine storage and regulated secretion. Loss of HID-1 causes a reduction in the number of LDCVs and affects their morphology and biochemical properties, due to impaired cargo sorting and dense core formation. HID-1 KO cells also exhibit defects in TGN acidification together with mislocalization of the Golgi-enriched vacuolar H+-ATPase subunit isoform a2. We propose that HID-1 influences early steps in LDCV formation by controlling dense core formation at the TGN.
AB - Large dense core vesicles (LDCVs) mediate the regulated release of neuropeptides and peptide hormones. They form at the trans-Golgi network (TGN), where their soluble content aggregates to form a dense core, but the mechanisms controlling biogenesis are still not completely understood. Recent studies have implicated the peripheral membrane protein HID-1 in neuropeptide sorting and insulin secretion. Using CRISPR/Cas9, we generated HID-1 KO rat neuroendocrine cells, and we show that the absence of HID-1 results in specific defects in peptide hormone and monoamine storage and regulated secretion. Loss of HID-1 causes a reduction in the number of LDCVs and affects their morphology and biochemical properties, due to impaired cargo sorting and dense core formation. HID-1 KO cells also exhibit defects in TGN acidification together with mislocalization of the Golgi-enriched vacuolar H+-ATPase subunit isoform a2. We propose that HID-1 influences early steps in LDCV formation by controlling dense core formation at the TGN.
UR - http://www.scopus.com/inward/record.url?scp=85038445847&partnerID=8YFLogxK
U2 - 10.1091/mbc.E17-08-0491
DO - 10.1091/mbc.E17-08-0491
M3 - Article
C2 - 29074564
AN - SCOPUS:85038445847
SN - 1059-1524
VL - 28
SP - 3870
EP - 3880
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
IS - 26
ER -