Journal article
Molecular basis of CRX/DNA recognition and stoichiometry at the Ret4 response element
Structure (London), Vol.32(10), pp.1751-1759.e4
10/2024
DOI: 10.1016/j.str.2024.07.004
PMCID: PMC11455607
PMID: 39084215
Abstract
Two retinal transcription factors, cone-rod homeobox (CRX) and neural retina leucine zipper (NRL), cooperate functionally and physically to control photoreceptor development and homeostasis. Mutations in CRX and NRL cause severe retinal diseases. Despite the roles of NRL and CRX, insight into their functions at the molecular level is lacking. Here, we have solved the crystal structure of the CRX homeodomain in complex with its cognate response element (Ret4) from the rhodopsin proximal promoter region. The structure reveals an unexpected 2:1 stoichiometry of CRX/Ret4 and unique orientation of CRX molecules on DNA, and it explains the mechanisms of pathogenic mutations in CRX. Mutations R41Q and E42K disrupt the CRX protein-protein contacts based on the structure and reduce the CRX/Ret4 binding stoichiometry, suggesting a novel disease mechanism. Furthermore, we show that NRL alters the stoichiometry and increases affinity of CRX binding at the rhodopsin promoter, which may enhance transcription of rod-specific genes and suppress transcription of cone-specific genes.
Details
- Title: Subtitle
- Molecular basis of CRX/DNA recognition and stoichiometry at the Ret4 response element
- Creators
- Dhiraj Srivastava - University of Iowa, Molecular Physiology and BiophysicsPavithra Gowribidanur-Chinnaswamy - University of Iowa, Molecular Physiology and BiophysicsParas Gaur - University of IowaMaria Spies - University of IowaAnand Swaroop - National Institutes of HealthNikolai O. Artemyev - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Structure (London), Vol.32(10), pp.1751-1759.e4
- DOI
- 10.1016/j.str.2024.07.004
- PMID
- 39084215
- PMCID
- PMC11455607
- NLM abbreviation
- Structure
- ISSN
- 0969-2126
- eISSN
- 1878-4186
- Publisher
- CELL PRESS; CAMBRIDGE
- Grant note
- US Depart-ment of Energy (DOE) Office of Science User Facility: DE-AC02-06CH11357, P30 GM138395
This work was supported by the National Institutes of Health grant RO1 EY-03445510843 to N.O.A. and by the National Eye Institute Intramural Research Program grant Z01EY000450 to A.S. D.S. was supported by Pediatric Ophthalmology Career Starter Research Grant from the Knights Templar Eye Foundation. We would like to acknowledge the use of resources at the Carver College of Medicine's Protein and Crystallography Facility at the University of Iowa. We thank Srinivas Chakravarty (BioCAT facility, Advanced Photon Source, now at Cytiva) and Jesse Hopkins (BioCAT facility, Advanced Photon Source) for help in SEC-MALS-SAXS and SEC-MALS data collection. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract no. DE-AC02- 06CH11357. BioCAT was supported by grant P30 GM138395 from the National Institute of General Medical Sciences of the National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily reflect the official views of the National Institute of General Medical Sciences or the National Institutes of Health.
- Language
- English
- Electronic publication date
- 07/2024
- Date published
- 10/2024
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Radiation Oncology; Biochemistry and Molecular Biology; Ophthalmology and Visual Sciences
- Record Identifier
- 9984691559202771
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