The vacuolar-ATPase complex and assembly factors, TMEM199 and CCDC115, control HIF1α prolyl hydroxylation by regulating cellular iron levels

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Abstract

Hypoxia Inducible transcription Factors (HIFs) are principally regulated by the 2-oxoglutarate and Iron(II) prolyl hydroxylase (PHD) enzymes, which hydroxylate the HIFα subunit, facilitating its proteasome-mediated degradation. Observations that HIFα hydroxylation can be impaired even when oxygen is sufficient emphasise the importance of understanding the complex nature of PHD regulation. Here, we use an unbiased genome-wide genetic screen in near-haploid human cells to uncover cellular processes that regulate HIF1α. We identify that genetic disruption of the Vacuolar H+ ATPase (V-ATPase), the key proton pump for endo-lysosomal acidification, and two previously uncharacterised V-ATPase assembly factors, TMEM199 and CCDC115, stabilise HIF1α in aerobic conditions. Rather than preventing the lysosomal degradation of HIF1α, disrupting the V-ATPase results in intracellular iron depletion, thereby impairing PHD activity and leading to HIF activation. Iron supplementation directly restores PHD catalytic activity following V-ATPase inhibition, revealing important links between the V-ATPase, iron metabolism and HIFs.

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Anna L Miles

Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom

Competing interests
The authors declare that no competing interests exist.
Stephen P Burr

Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom

Competing interests
The authors declare that no competing interests exist.
Guinevere L Grice

Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom

Competing interests
The authors declare that no competing interests exist.
James A Nathan

Cambridge Institute for Medical Research, Department of Medicine, University of Cambridge, Cambridge, United Kingdom

For correspondence
jan33@cam.ac.uk

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-0248-1632

Funding

Wellcome (102770/Z/13/Z)

Stephen P Burr
Guinevere L Grice
James A Nathan

Medical Research Council (MR/K50127X/1)

Anna L Miles
James A Nathan

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

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This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.