{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T23:35:06Z","timestamp":1785800106876,"version":"3.56.0"},"reference-count":49,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2024,11,20]],"date-time":"2024-11-20T00:00:00Z","timestamp":1732060800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pathogenic variants in KCNC3, which encodes the voltage-gated potassium channel Kv3.3, are associated with spinocerebellar ataxia type 13. SCA13 is a neurodegenerative disease characterized by ataxia, dysarthria and oculomotor dysfunction, often in combination with other signs and symptoms such as cognitive impairment. Known disease-causing variants are localized in the protein coding regions and predominantly in the transmembrane and voltage sensing domains. In a patient with an ataxic movement disorder and progressive cognitive decline, the c.-6C&gt;A variant was detected in the Kozak sequence of KCNC3. The Kozak sequence is responsible for efficient initiation of translation. Functional analysis of the new c.-6C&gt;A variant and the upstream 5\u2019-UTR region of KCNC3 by luciferase assays, quantitative PCR and methylation analysis shows increased protein expression but no effect on transcription rate. Therefore, increased translation initiation of KCNC3 transcripts compared to wild-type Kozak sequence seems to be the cause of the increased expression. Variants in the regulatory elements of disease-causing genes probably play an underestimated role.<\/jats:p>","DOI":"10.3390\/ijms252212444","type":"journal-article","created":{"date-parts":[[2024,11,20]],"date-time":"2024-11-20T03:57:04Z","timestamp":1732075024000},"page":"12444","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Kv3.3 Expression Enhanced by a Novel Variant in the Kozak Sequence of KCNC3"],"prefix":"10.3390","volume":"25","author":[{"given":"Marlen Colleen","family":"Reis","sequence":"first","affiliation":[{"name":"Institute of Human Genetics, Department of Medicine, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frauke","family":"H\u00e4rtel","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Department of Medicine, Justus Liebig University Giessen, 35390 Giessen, Germany"},{"name":"Institute of Physiology, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9392-1495","authenticated-orcid":false,"given":"Antje Maria","family":"Richter","sequence":"additional","affiliation":[{"name":"Institute of Genetics, Department of Biology, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michaela","family":"Wei\u00df","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Department of Medicine, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lea-Theresa","family":"M\u00f6sle","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Department of Medicine, Justus Liebig University Giessen, 35390 Giessen, Germany"},{"name":"Innere Medizinische Klinik II, Klinikum Memmingen, 87700 Memmingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6407-1236","authenticated-orcid":false,"given":"Reinhard Heinrich","family":"Dammann","sequence":"additional","affiliation":[{"name":"Institute of Genetics, Department of Biology, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dagmar","family":"Nolte","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Department of Medicine, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Roos, D., and de Boer, M. (2021). Mutations in cis that affect mRNA synthesis, processing and translation. Biochim. Biophys. Acta Mol. Basis Dis., 1867.","DOI":"10.1016\/j.bbadis.2021.166166"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1007\/s00018-002-8420-z","article-title":"DNA methylation and the regulation of gene transcription","volume":"59","author":"Attwood","year":"2002","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1038\/nrg.2016.83","article-title":"Transcription factors as readers and effectors of DNA methylation","volume":"17","author":"Zhu","year":"2016","journal-title":"Nat. Rev. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"8125","DOI":"10.1093\/nar\/15.20.8125","article-title":"An analysis of 5\u2032-noncoding sequences from 699 vertebrate messenger RNAs","volume":"15","author":"Kozak","year":"1987","journal-title":"Nucleic Acids Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1016\/j.tibs.2019.07.001","article-title":"Conservation and Variability of the AUG Initiation Codon Context in Eukaryotes","volume":"44","author":"Osnaya","year":"2019","journal-title":"Trends Biochem. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1007\/s00335-002-4002-5","article-title":"Emerging links between initiation of translation and human diseases","volume":"13","author":"Kozak","year":"2002","journal-title":"Mamm. Genome"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Nicolle, R., Altin, N., Siquier-Pernet, K., Salignac, S., Blanc, P., Munnich, A., Bole-Feysot, C., Malan, V., Caron, B., and Nitschk\u00e9, P. (2023). A non-coding variant in the Kozak sequence of RARS2 strongly decreases protein levels and causes pontocerebellar hypoplasia. BMC Med. Genom., 16.","DOI":"10.1186\/s12920-023-01582-z"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1182\/blood.V94.1.186.413k19_186_191","article-title":"Kozak sequence polymorphism of the glycoprotein (GP) Ibalpha gene is a major determinant of the plasma membrane levels of the platelet GP Ib-IX-V complex","volume":"94","author":"Li","year":"1999","journal-title":"Blood"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2684","DOI":"10.1210\/en.2004-1617","article-title":"A Graves\u2019 disease-associated Kozak sequence single-nucleotide polymorphism enhances the efficiency of CD40 gene translation: A case for translational pathophysiology","volume":"146","author":"Jacobson","year":"2005","journal-title":"Endocrinology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"112229","DOI":"10.1016\/j.yexcr.2020.112229","article-title":"Some like it translated: Small ORFs in the 5\u2032UTR","volume":"396","author":"Renz","year":"2020","journal-title":"Exp. Cell Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/s41572-019-0074-3","article-title":"Spinocerebellar ataxia","volume":"5","author":"Klockgether","year":"2019","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_12","unstructured":"Adam, M.P., Feldman, J., Mirzaa, G.M., Pagon, R.A., Wallace, S.E., and Amemiya, A. (2024, October 25). Spinocerebellar Ataxia Type 13, GeneReviews\u00ae [Internet], Available online: https:\/\/www.ncbi.nlm.nih.gov\/books\/NBK1225\/."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1038\/ng1758","article-title":"Mutations in voltage-gated potassium channel KCNC3 cause degenerative and developmental central nervous system phenotypes","volume":"38","author":"Waters","year":"2006","journal-title":"Nat. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1016\/0888-7543(92)90365-Y","article-title":"Genomic organization, nucleotide sequence, and cellular distribution of a Shaw-related potassium channel gene, Kv3.3, and mapping of Kv3.3 and Kv3.4 to human chromosomes 19 and 1","volume":"12","author":"Ghanshani","year":"1992","journal-title":"Genomics"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1002\/humu.21165","article-title":"KCNC3: Phenotype, mutations, channel biophysics-a study of 260 familial ataxia patients","volume":"31","author":"Figueroa","year":"2010","journal-title":"Hum. Mutat."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3106","DOI":"10.1093\/brain\/awt236","article-title":"Next generation sequencing for molecular diagnosis of neurological disorders using ataxias as a model","volume":"136","author":"Kwasniewska","year":"2013","journal-title":"Brain"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Duarri, A., Nibbeling, E.A., Fokkens, M.R., Meijer, M., Boerrigter, M., Verschuuren-Bemelmans, C.C., Kremer, B.P., van de Warrenburg, B.P., Dooijes, D., and Boddeke, E. (2015). Functional analysis helps to define KCNC3 mutational spectrum in Dutch ataxia cases. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0116599"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1817","DOI":"10.1093\/brain\/awv117","article-title":"De novo point mutations in patients diagnosed with ataxic cerebral palsy","volume":"138","author":"Perkins","year":"2015","journal-title":"Brain"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1016\/j.cell.2016.02.009","article-title":"Kv3.3 Channels Bind Hax-1 and Arp2\/3 to Assemble a Stable Local Actin Network that Regulates Channel Gating","volume":"165","author":"Zhang","year":"2016","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Khare, S., Nick, J.A., Zhang, Y., Galeano, K., Butler, B., Khoshbouei, H., Rayaprolu, S., Hathorn, T., Ranum, L.P.W., and Smithson, L. (2017). A KCNC3 mutation causes a neurodevelopmental, non-progressive SCA13 subtype associated with dominant negative effects and aberrant EGFR trafficking. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0173565"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1007\/s12311-018-0950-5","article-title":"C-terminal proline deletions in KCNC3 cause delayed channel inactivation and an adult-onset progressive SCA13 with spasticity","volume":"17","author":"Khare","year":"2018","journal-title":"Cerebellum"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1038\/s10038-019-0717-y","article-title":"Genetic screening for potassium channel mutations in Japanese autosomal dominant spinocerebellar ataxia","volume":"65","author":"Tada","year":"2020","journal-title":"J. Hum. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1016\/j.ajhg.2023.05.009","article-title":"Extreme phenotypic heterogeneity in non-expansion spinocerebellar ataxias","volume":"110","author":"Cunha","year":"2023","journal-title":"Am. J. Hum. Genet."},{"key":"ref_24","unstructured":"Phan, L., Jin, Y., Zhang, H., Qiang, W., Shekhtman, E., Shao, D., Revoe, D., Villamarin, R., Ivanchenko, E., and Kimura, M. (2024, November 11). ALFA: Allele Frequency Aggregator. National Center for Biotechnology Information, U.S. National Library of Medicine 2020, Available online: www.ncbi.nlm.nih.gov\/snp\/docs\/gsr\/alfa\/."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1038\/nmeth.2890","article-title":"MutationTaster2: Mutation prediction for the deep-sequencing age","volume":"11","author":"Schwarz","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"D1143","DOI":"10.1093\/nar\/gkad989","article-title":"CADD v1.7: Using protein language models, regulatory CNNs and other nucleotide-level scores to improve genome-wide variant predictions","volume":"52","author":"Schubach","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1038\/ng1728","article-title":"Spectrin mutations cause spinocerebellar ataxia type 5","volume":"38","author":"Ikeda","year":"2006","journal-title":"Nat. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Lise, S., Clarkson, Y., Perkins, E., Kwasniewska, A., Sadighi Akha, E., Parolin Schnekenberg, R., Suminaite, D., Hope, J., Baker, I., and Gregory, L. (2012). Recessive mutations in SPTBN2 implicate \u03b2-III spectrin in both cognitive and motor development. PLoS Genet., 8.","DOI":"10.1371\/journal.pgen.1003074"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and guidelines for the interpretation of sequence var-iants: A joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"ref_30","first-page":"226","article-title":"Neural network prediction of translation initiation sites in eukaryotes: Perspectives for EST and genome analysis","volume":"5","author":"Pedersen","year":"1997","journal-title":"Proc. Int. Conf. Intell. Syst. Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Reuter, K., Biehl, A., Koch, L., and Helms, V. (2016). PreTIS: A Tool to Predict Non-canonical 5\u2032 UTR Translational Initiation Sites in Human and Mouse. PLoS Comput. Biol., 12.","DOI":"10.1371\/journal.pcbi.1005170"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1523\/JNEUROSCI.1132-21.2021","article-title":"Ankyrin-R Links Kv3.3 to the Spectrin Cytoskeleton and Is Required for Purkinje Neuron Survival","volume":"42","author":"Stevens","year":"2022","journal-title":"J. Neurosci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1042\/BJ20130034","article-title":"Spinocerebellar ataxia-13 Kv3.3 potassium channels: Arginine-to-histidine mutations affect both functional and protein expression on the cell surface","volume":"454","author":"Zhao","year":"2013","journal-title":"Biochem. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e57358","DOI":"10.7554\/eLife.57358","article-title":"Infant and adult SCA13 mutations differentially affect Purkinje cell excitability, maturation, and viability in vivo","volume":"9","author":"Hsieh","year":"2020","journal-title":"eLife"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1073\/pnas.81.4.1154","article-title":"beta-Thalassemia in American Blacks: Novel mutations in the \u201cTATA\u201d box and an acceptor splice site","volume":"81","author":"Antonarakis","year":"1984","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"828","DOI":"10.1111\/j.1365-2141.2004.04835.x","article-title":"Thalassaemia mutations within the 5\u2032UTR of the human beta-globin gene disrupt transcription","volume":"124","author":"Sgourou","year":"2004","journal-title":"Br. J. Haematol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"936","DOI":"10.1086\/504044","article-title":"Promoter mutations that increase amyloid precursor-protein expression are associated with Alzheimer disease","volume":"78","author":"Theuns","year":"2006","journal-title":"Am. J. Hum. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ghorbani, F., de Boer, E.N., Fokkens, M.R., de Boer-Bergsma, J., Verschuuren-Bemelmans, C.C., Wierenga, E., Kasaei, H., Noordermeer, D., Verbeek, D.S., and Westers, H. (2024). Identification and Copy Number Variant Analysis of Enhancer Regions of Genes Causing Spinocerebellar Ataxia. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252011205"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/j.bbrc.2010.01.002","article-title":"Screening of Kozak-motif-located SNPs and analysis of their association with human diseases","volume":"392","author":"Xu","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2732","DOI":"10.1002\/ajmg.a.36703","article-title":"A mutation in the Kozak sequence of GATA4 hampers translation in a family with atrial septal defects","volume":"164A","author":"Mohan","year":"2014","journal-title":"Am. J. Med. Genet. A."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"10756","DOI":"10.1093\/nar\/gkac799","article-title":"Translational enhancement by base editing of the Kozak sequence rescues haploinsufficiency","volume":"50","author":"Ambrosini","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10075","DOI":"10.1093\/nar\/gkad687","article-title":"Precise genome editing of the Kozak sequence enables bidirectional and quantitative modulation of protein translation toanticipated levels without affecting transcription","volume":"51","author":"Xie","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/1750-1172-9-38","article-title":"PMP22 related neuropathies: Charcot-Marie-Tooth disease type 1a and Hereditary Neuropathy with liability to Pressure Palsies","volume":"9","author":"Verhamme","year":"2014","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1016\/j.hrthm.2011.02.021","article-title":"Transient outward current (I(to)) gain-of-function mutations in the KCND3-encoded Kv4.3 potassium channel and Brugada syndrome","volume":"8","author":"Giudicessi","year":"2011","journal-title":"Heart Rhythm."},{"key":"ref_45","first-page":"e34597","article-title":"Association of Cardiac Electrical Disorders with KCND3 Gene Mutation","volume":"15","author":"Ahammed","year":"2023","journal-title":"Cureus"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1002\/humu.24384","article-title":"uORF-introducing variants in the 5\u2032UTR of the NIPBL gene as a cause of Cornelia de Lange syndrome","volume":"43","author":"Coursimault","year":"2022","journal-title":"Hum. Mutat."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1552","DOI":"10.1007\/s12311-023-01629-y","article-title":"Diagnostic Yield of NGS Tests for Hereditary Ataxia: A Systematic Review","volume":"23","author":"Tenorio","year":"2023","journal-title":"Cerebellum"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Reis, M.C., Patrun, J., Ackl, N., Winter, P., Scheifele, M., Danek, A., and Nolte, D. (2022). A Severe Dementia Syndrome Caused by Intron Retention and Cryptic Splice Site Activation in STUB1 and Exacerbated by TBP Repeat Expansions. Front. Mol. Neurosci., 15.","DOI":"10.3389\/fnmol.2022.878236"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1038\/s41586-020-2308-7","article-title":"The mutational constraint spectrum quantified from variation in 141,456 humans","volume":"581","author":"Karczewski","year":"2020","journal-title":"Nature"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/22\/12444\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:35:33Z","timestamp":1760114133000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/22\/12444"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,20]]},"references-count":49,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2024,11]]}},"alternative-id":["ijms252212444"],"URL":"https:\/\/doi.org\/10.3390\/ijms252212444","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,20]]}}}