{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T22:33:40Z","timestamp":1768516420186,"version":"3.49.0"},"posted":{"date-parts":[[2024,9,19]]},"group-title":"Biochemistry","reference-count":57,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,9,19]],"date-time":"2024-09-19T00:00:00Z","timestamp":1726704000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,9,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Spinocerebellar ataxia type 5 (SCA5) mutations in the protein \u03b2-III-spectrin cluster to the N-terminal actin-binding domain (ABD) and the central spectrin-repeat domains (SRDs). We previously reported that a common molecular consequence of ABD-localized SCA5 mutations is increased actin binding. However, little is known about the molecular consequences of the SRD-localized mutations. It is known that the SRDs of \u03b2-spectrin proteins interact with \u03b1-spectrin to form an \u03b1\/\u03b2-spectrin dimer. In addition, it is known that SRDs neighbouring the \u03b2-spectrin ABD enhance actin binding. Here, we tested the impact of the SRD-localized R480W and the E532_M544del mutations on the binding of \u03b2-III-spectrin to \u03b1-II-spectrin and actin. Using multiple experimental approaches, we show that both the R480W and E532_M544del mutants can bind \u03b1-II-spectrin. However, E532_M544del causes partial uncoupling of complementary SRDs in the \u03b1\/\u03b2-spectrin dimer. Further, the R480W mutant forms large intracellular inclusions when co-expressed with \u03b1-II-spectrin in cells, supporting that R480W mutation grossly disrupts the \u03b1-II\/\u03b2-III-spectrin physical complex. Moreover, actin-binding assays show that E532_M544del, but not R480W, increases \u03b2-III-spectrin actin binding. Altogether, these data support that SRD-localized mutations alter key interactions of \u03b2-III-spectrin with \u03b1-II-spectrin and actin.<\/jats:p>","DOI":"10.1101\/2024.09.17.613313","type":"posted-content","created":{"date-parts":[[2024,9,20]],"date-time":"2024-09-20T00:25:34Z","timestamp":1726791934000},"source":"Crossref","is-referenced-by-count":0,"title":["Molecular consequences of SCA5 mutations in the spectrin-repeat domains of \u03b2-III-spectrin"],"prefix":"10.64898","author":[{"given":"Sarah A.","family":"Denha","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naomi R.","family":"DeLaet","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abeer W.","family":"Abukamil","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angelica 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