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Biochemical defects in ABCR protein variants associated with human retinopathies

Nature Genetics volume 26, pages 242–246 (2000)Cite this article

Abstract

Mutations in the gene encoding ABCR (ABCA4), a photoreceptor-specific ATP-binding cassette (ABC) transporter1,2,3,4, are responsible for autosomal recessive Stargardt disease (STGD), an early onset macular degeneration1,5,6,7,8,9,10,11, and some forms of autosomal recessive cone-rod dystrophy12 and autosomal recessive retinitis pigmentosa12,13,14. Heterozygosity for ABCA4 mutations may also represent a risk factor for age-related macular degeneration15 (AMD), although this idea is controversial7,16,17. An ongoing challenge in the analysis of ABCA4-based retinopathies arises from the observation that most of the ABCA4 sequence variants identified so far are missense mutations that are rare in both patient and control populations. With the current sample size of most sequence variants, one cannot determine statistically whether a particular sequence variant is pathogenic or neutral. A related challenge is to determine the degree to which each pathogenic variant impairs ABCR function, as genotype-phenotype analyses indicate that age of onset and disease severity correlate with different ABCA4 alleles6,8,9,10. To address these questions, we performed a functional analysis of human ABCR and its variants. These experiments reveal a wide spectrum of biochemical defects in these variants and provide insight into the transport mechanism of ABCR.

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Figure 1: Production of functional ABCR in transiently transfected 293 cells.
Figure 2: Locations of 37 naturally occurring ABCR sequence variants and 4 synthetic mutations.
Figure 3: Protein yield and ATP-binding capacity of 37 naturally occurring ABCR variants produced in transiently transfected 293 cells.
Figure 4: Effect of retinal on ATP hydrolysis by a subset of naturally occurring ABCR variants.
Figure 5: Effect of retinal on ATP hydrolysis by naturally occurring and synthetic mutants in the NBDs.

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Acknowledgements

We thank C. Riley, C. Davenport, M. Kazienko and J. Ptak for DNA synthesis and sequencing; R. Molday for the gift of cells producing Rim3F4; R. Allikmets, J. Lupski and E. Stone for sharing their data on ABCR variants and for helpful discussions; S. Almashanu for advice; and A. Rattner and P. Tong for helpful comments on the manuscript. This work was supported by the Howard Hughes Medical Institute and the National Eye Institute (NIH).

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Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Hui Sun, Philip M. Smallwood & Jeremy Nathans

  2. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Jeremy Nathans

  3. Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Jeremy Nathans

  4. Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Hui Sun, Philip M. Smallwood & Jeremy Nathans

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Correspondence to Jeremy Nathans.

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Sun, H., Smallwood, P. & Nathans, J. Biochemical defects in ABCR protein variants associated with human retinopathies. Nat Genet 26, 242–246 (2000). https://doi.org/10.1038/79994

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