TY - JOUR
T1 - Comprehensive identification of mRNA isoforms reveals the diversity of neural cell-surface molecules with roles in retinal development and disease
AU - Ray, Thomas A.
AU - Cochran, Kelly
AU - Kozlowski, Chris
AU - Wang, Jingjing
AU - Alexander, Graham
AU - Cady, Martha A.
AU - Spencer, William J.
AU - Ruzycki, Philip A.
AU - Clark, Brian S.
AU - Laeremans, Annelies
AU - He, Ming Xiao
AU - Wang, Xiaoming
AU - Park, Emily
AU - Hao, Ying
AU - Iannaccone, Alessandro
AU - Hu, Gary
AU - Fedrigo, Olivier
AU - Skiba, Nikolai P.
AU - Arshavsky, Vadim Y.
AU - Kay, Jeremy N.
N1 - Funding Information:
This work was supported by the National Eye Institute (EY024694 and EY030611 to J.N. K., EY026344 to T.A.R., EY5722 to Duke University, EY027844 to B.S.C.); Pew Charitable Trusts (J.N.K.); McKnight Endowment Fund for Neuroscience (J.N.K.); a Duke University School of Medicine Kahn Neurotechnology Award (J.N.K.); Foundation Fighting Blindness (Individual Investigator Award BR‐CMM‐0619‐0767‐DUKE to J.N.K.); and Research to Prevent Blindness (Unrestricted Grant to Duke University; Unrestricted Grant to Washington University). Duke Research Computing provided computing resources for data analysis. This project benefitted from data storage capacity provided by the Duke Data Commons, a data storage installation underwritten by the NIH (1S10OD018164-01). We thank the Duke Transgenic and Knockout Mouse Shared Resource for their assistance in making mutant mice, Liz Tseng (PacBio) for assistance with Iso-Seq 3, and Ariane Pereira for mouse colony management.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Genes encoding cell-surface proteins control nervous system development and are implicated in neurological disorders. These genes produce alternative mRNA isoforms which remain poorly characterized, impeding understanding of how disease-associated mutations cause pathology. Here we introduce a strategy to define complete portfolios of full-length isoforms encoded by individual genes. Applying this approach to neural cell-surface molecules, we identify thousands of unannotated isoforms expressed in retina and brain. By mass spectrometry we confirm expression of newly-discovered proteins on the cell surface in vivo. Remarkably, we discover that the major isoform of a retinal degeneration gene, CRB1, was previously overlooked. This CRB1 isoform is the only one expressed by photoreceptors, the affected cells in CRB1 disease. Using mouse mutants, we identify a function for this isoform at photoreceptor-glial junctions and demonstrate that loss of this isoform accelerates photoreceptor death. Therefore, our isoform identification strategy enables discovery of new gene functions relevant to disease.
AB - Genes encoding cell-surface proteins control nervous system development and are implicated in neurological disorders. These genes produce alternative mRNA isoforms which remain poorly characterized, impeding understanding of how disease-associated mutations cause pathology. Here we introduce a strategy to define complete portfolios of full-length isoforms encoded by individual genes. Applying this approach to neural cell-surface molecules, we identify thousands of unannotated isoforms expressed in retina and brain. By mass spectrometry we confirm expression of newly-discovered proteins on the cell surface in vivo. Remarkably, we discover that the major isoform of a retinal degeneration gene, CRB1, was previously overlooked. This CRB1 isoform is the only one expressed by photoreceptors, the affected cells in CRB1 disease. Using mouse mutants, we identify a function for this isoform at photoreceptor-glial junctions and demonstrate that loss of this isoform accelerates photoreceptor death. Therefore, our isoform identification strategy enables discovery of new gene functions relevant to disease.
UR - http://www.scopus.com/inward/record.url?scp=85087395808&partnerID=8YFLogxK
U2 - 10.1038/s41467-020-17009-7
DO - 10.1038/s41467-020-17009-7
M3 - Article
C2 - 32620864
AN - SCOPUS:85087395808
SN - 2041-1723
VL - 11
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 3328
ER -