Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane

John J. Castorino, Shannon M. Gallagher-Colombo, Alex V. Levin, Paul G FitzGerald, Jessica Polishook, Barbara Kloeckener-Gruissem, Eric Ostertag, Nancy J. Philp

Research output: Contribution to journalArticle

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Abstract

Purpose. SLC16A12 encodes an orphan member of the monocarboxylate transporter family, MCT12. A nonsense mutation in SLC16A12 (c.643C>T; p.Q215X) causes juvenile cataract with a dominant inheritance pattern. In the present study, in vitro and in vivo experimental models were used to gain insight into how the SLC16A12 (c.643C>T) mutation leads to cataract formation. Methods. MCT12 peptide antibodies were generated and used to examine the expression of MCT12 in the lens using immuno-confocal microscopy. To determine whether loss of Slc16a12 resulted in cataract formation, a Slc16a12 hypomorphic rat generated by transposon insertional mutagenesis was characterized using RT-PCR, slit lamp microscopy and histologic methods. Exogenous expression of MCT12 and MCT12:214Δ, a mimic of the mutant allele, were used to assess protein expression and trafficking. Results. MCT12 protein was detected in the lens epithelium and secondary fiber cells at postnatal day 1. In the Slc16a12TKO rat, complete loss of MCT12 did not result in any detectable ocular phenotype. Exogenous expression of MCT12-GFP and MCT12:214Δ-GFP revealed that the full-length protein was trafficked to the plasma membrane (PM), whereas the truncated protein was retained in the endoplasmic reticulum (ER). When both MCT12 and MCT12:214Δ were coexpressed, to mimic the heterozygous patient genotype, the truncated protein was retained in the ER whereas full-length MCT12 was trafficked to the PM. Furthermore, MCT12 was identified as another MCT isoform that requires CD147 for trafficking to the cell surface. Conclusions. These data support a model whereby the SLC16A12 (c.643C>T) mutation causes juvenile cataract by a defect in protein trafficking rather than by haploinsufficiency.

Original languageEnglish (US)
Pages (from-to)6774-6784
Number of pages11
JournalInvestigative Ophthalmology and Visual Science
Volume52
Issue number9
DOIs
StatePublished - Aug 2011

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Protein Transport
Cataract
Cell Membrane
Mutation
Endoplasmic Reticulum
Lenses
Proteins
Haploinsufficiency
Inheritance Patterns
Orphaned Children
Nonsense Codon
Insertional Mutagenesis
Confocal Microscopy
Protein Isoforms
Theoretical Models
Epithelium
Alleles
Genotype
Phenotype
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane. / Castorino, John J.; Gallagher-Colombo, Shannon M.; Levin, Alex V.; FitzGerald, Paul G; Polishook, Jessica; Kloeckener-Gruissem, Barbara; Ostertag, Eric; Philp, Nancy J.

In: Investigative Ophthalmology and Visual Science, Vol. 52, No. 9, 08.2011, p. 6774-6784.

Research output: Contribution to journalArticle

Castorino, JJ, Gallagher-Colombo, SM, Levin, AV, FitzGerald, PG, Polishook, J, Kloeckener-Gruissem, B, Ostertag, E & Philp, NJ 2011, 'Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane', Investigative Ophthalmology and Visual Science, vol. 52, no. 9, pp. 6774-6784. https://doi.org/10.1167/iovs.10-6579
Castorino, John J. ; Gallagher-Colombo, Shannon M. ; Levin, Alex V. ; FitzGerald, Paul G ; Polishook, Jessica ; Kloeckener-Gruissem, Barbara ; Ostertag, Eric ; Philp, Nancy J. / Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane. In: Investigative Ophthalmology and Visual Science. 2011 ; Vol. 52, No. 9. pp. 6774-6784.
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abstract = "Purpose. SLC16A12 encodes an orphan member of the monocarboxylate transporter family, MCT12. A nonsense mutation in SLC16A12 (c.643C>T; p.Q215X) causes juvenile cataract with a dominant inheritance pattern. In the present study, in vitro and in vivo experimental models were used to gain insight into how the SLC16A12 (c.643C>T) mutation leads to cataract formation. Methods. MCT12 peptide antibodies were generated and used to examine the expression of MCT12 in the lens using immuno-confocal microscopy. To determine whether loss of Slc16a12 resulted in cataract formation, a Slc16a12 hypomorphic rat generated by transposon insertional mutagenesis was characterized using RT-PCR, slit lamp microscopy and histologic methods. Exogenous expression of MCT12 and MCT12:214Δ, a mimic of the mutant allele, were used to assess protein expression and trafficking. Results. MCT12 protein was detected in the lens epithelium and secondary fiber cells at postnatal day 1. In the Slc16a12TKO rat, complete loss of MCT12 did not result in any detectable ocular phenotype. Exogenous expression of MCT12-GFP and MCT12:214Δ-GFP revealed that the full-length protein was trafficked to the plasma membrane (PM), whereas the truncated protein was retained in the endoplasmic reticulum (ER). When both MCT12 and MCT12:214Δ were coexpressed, to mimic the heterozygous patient genotype, the truncated protein was retained in the ER whereas full-length MCT12 was trafficked to the PM. Furthermore, MCT12 was identified as another MCT isoform that requires CD147 for trafficking to the cell surface. Conclusions. These data support a model whereby the SLC16A12 (c.643C>T) mutation causes juvenile cataract by a defect in protein trafficking rather than by haploinsufficiency.",
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T1 - Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane

AU - Castorino, John J.

AU - Gallagher-Colombo, Shannon M.

AU - Levin, Alex V.

AU - FitzGerald, Paul G

AU - Polishook, Jessica

AU - Kloeckener-Gruissem, Barbara

AU - Ostertag, Eric

AU - Philp, Nancy J.

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N2 - Purpose. SLC16A12 encodes an orphan member of the monocarboxylate transporter family, MCT12. A nonsense mutation in SLC16A12 (c.643C>T; p.Q215X) causes juvenile cataract with a dominant inheritance pattern. In the present study, in vitro and in vivo experimental models were used to gain insight into how the SLC16A12 (c.643C>T) mutation leads to cataract formation. Methods. MCT12 peptide antibodies were generated and used to examine the expression of MCT12 in the lens using immuno-confocal microscopy. To determine whether loss of Slc16a12 resulted in cataract formation, a Slc16a12 hypomorphic rat generated by transposon insertional mutagenesis was characterized using RT-PCR, slit lamp microscopy and histologic methods. Exogenous expression of MCT12 and MCT12:214Δ, a mimic of the mutant allele, were used to assess protein expression and trafficking. Results. MCT12 protein was detected in the lens epithelium and secondary fiber cells at postnatal day 1. In the Slc16a12TKO rat, complete loss of MCT12 did not result in any detectable ocular phenotype. Exogenous expression of MCT12-GFP and MCT12:214Δ-GFP revealed that the full-length protein was trafficked to the plasma membrane (PM), whereas the truncated protein was retained in the endoplasmic reticulum (ER). When both MCT12 and MCT12:214Δ were coexpressed, to mimic the heterozygous patient genotype, the truncated protein was retained in the ER whereas full-length MCT12 was trafficked to the PM. Furthermore, MCT12 was identified as another MCT isoform that requires CD147 for trafficking to the cell surface. Conclusions. These data support a model whereby the SLC16A12 (c.643C>T) mutation causes juvenile cataract by a defect in protein trafficking rather than by haploinsufficiency.

AB - Purpose. SLC16A12 encodes an orphan member of the monocarboxylate transporter family, MCT12. A nonsense mutation in SLC16A12 (c.643C>T; p.Q215X) causes juvenile cataract with a dominant inheritance pattern. In the present study, in vitro and in vivo experimental models were used to gain insight into how the SLC16A12 (c.643C>T) mutation leads to cataract formation. Methods. MCT12 peptide antibodies were generated and used to examine the expression of MCT12 in the lens using immuno-confocal microscopy. To determine whether loss of Slc16a12 resulted in cataract formation, a Slc16a12 hypomorphic rat generated by transposon insertional mutagenesis was characterized using RT-PCR, slit lamp microscopy and histologic methods. Exogenous expression of MCT12 and MCT12:214Δ, a mimic of the mutant allele, were used to assess protein expression and trafficking. Results. MCT12 protein was detected in the lens epithelium and secondary fiber cells at postnatal day 1. In the Slc16a12TKO rat, complete loss of MCT12 did not result in any detectable ocular phenotype. Exogenous expression of MCT12-GFP and MCT12:214Δ-GFP revealed that the full-length protein was trafficked to the plasma membrane (PM), whereas the truncated protein was retained in the endoplasmic reticulum (ER). When both MCT12 and MCT12:214Δ were coexpressed, to mimic the heterozygous patient genotype, the truncated protein was retained in the ER whereas full-length MCT12 was trafficked to the PM. Furthermore, MCT12 was identified as another MCT isoform that requires CD147 for trafficking to the cell surface. Conclusions. These data support a model whereby the SLC16A12 (c.643C>T) mutation causes juvenile cataract by a defect in protein trafficking rather than by haploinsufficiency.

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