Abstract
The human cis-prenyltransferase (hcis-PT) is an enzymatic complex essential for protein N-glycosylation. Synthesizing the precursor of the glycosyl carrier dolichol-phosphate, mutations in hcis-PT cause severe human diseases. Here, we reveal that hcis-PT exhibits a heterotetrameric assembly in solution, consisting of two catalytic dehydrodolichyl diphosphate synthase (DHDDS) and inactive Nogo-B receptor (NgBR) heterodimers. Importantly, the 2.3 Å crystal structure reveals that the tetramer assembles via the DHDDS C-termini as a dimer-of-heterodimers. Moreover, the distal C-terminus of NgBR transverses across the interface with DHDDS, directly participating in active-site formation and the functional coupling between the subunits. Finally, we explored the functional consequences of disease mutations clustered around the active-site, and in combination with molecular dynamics simulations, we propose a mechanism for hcis-PT dysfunction in retinitis pigmentosa. Together, our structure of the hcis-PT complex unveils the dolichol synthesis mechanism and its perturbation in disease.
| Original language | English |
|---|---|
| Article number | 5273 |
| Journal | Nature Communications |
| Volume | 11 |
| Issue number | 1 |
| DOIs | |
| State | Published - 19 Oct 2020 |
| Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2020, The Author(s).
Funding
We thank the staff of I03 at the Diamond Light Source for assistance with diffraction experimentation and Dr. Reuven Wiener for technical assistance with diffraction data collection. This work was performed in partial fulfillment of the requirements for a Ph.D. degree of M.L.B.-E., Sackler Faculty of Medicine, Tel-Aviv University, Israel. This work was supported by the Israel Science Foundation (grant 1721/16) (Y.H.), the Israel Cancer Research Foundation grants 01214 (Y.H.) and 19202 (M.G.), and from the German‐ Israeli Foundation for Scientific Research and Development (grant No. I‐2425‐418.13/ 2016) (Y.H.). Support also came from the I‐CORE Program of the Planning and Budgeting Committee and the Israel Science Foundation (grant 1775/12) (Y.H.), Recanati Foundation (M.G.), Marguerite Stolz Research Fellowship (Y.H.), Kahn foundation's Orion project, Tel Aviv Medical Center, Israel (M.G.), Israel Cancer Association (grant 20200037) (Y.H. and M.G.), and the Claire and Amedee Maratier Institute for the Study of Blindness and Visual Disorders, Sackler Faculty of Medicine, Tel-Aviv University (Y.H. and M.G.). Access to MS installation was funded by the EU Horizon 2020 grant EU_FT–ICR_MS project number 731077 and by CIISB LM2018127. P.M. and P.V. support from MEYS CZ funds CZ.1.05/1.1.00/02.0109 is gratefully acknowledged.
| Funders | Funder number |
|---|---|
| CIISB | LM2018127 |
| German‐ Israeli Foundation for Scientific Research and Development | 1775/12, I‐2425‐418.13/ 2016 |
| Israel Cancer Research Foundation | 19202, 01214 |
| Recanati Foundation | |
| Sackler Faculty of Medicine | |
| Visual Disorders | |
| Israel Cancer Research Fund | |
| Horizon 2020 Framework Programme | |
| German-Israeli Foundation for Scientific Research and Development | |
| Ministry of Education, Youth and Science | |
| Israel Cancer Association | 20200037 |
| Israel Science Foundation | 1721/16 |
| Claire and Amédée Maratier Institute for the Study of Blindness and Visual Disorders, Tel Aviv University | |
| Horizon 2020 | 731077 |
| Tel Aviv Sourasky Medical Center | |
| Sackler Faculty of Medicine, Tel-Aviv University |
