title: A new cytoplasmic interaction between junctin and ryanodine receptor Ca2+ release channels.
abstract: Junctin, a non-catalytic splice variant encoded by the aspartate-β-hydroxylase (Asph) gene, is inserted into the membrane of the sarcoplasmic reticulum (SR) Ca(2+) store where it modifies Ca(2+) signalling in the heart and skeletal muscle through its regulation of ryanodine receptor (RyR) Ca(2+) release channels. Junctin is required for normal muscle function as its knockout leads to abnormal Ca(2+) signalling, muscle dysfunction and cardiac arrhythmia. However, the nature of the molecular interaction between junctin and RyRs is largely unknown and was assumed to occur only in the SR lumen. We find that there is substantial binding of RyRs to full junctin, and the junctin luminal and, unexpectedly, cytoplasmic domains. Binding of these different junctin domains had distinct effects on RyR1 and RyR2 activity: full junctin in the luminal solution increased RyR channel activity by ∼threefold, the C-terminal luminal interaction inhibited RyR channel activity by ∼50%, and the N-terminal cytoplasmic binding produced an ∼fivefold increase in RyR activity. The cytoplasmic interaction between junctin and RyR is required for luminal binding to replicate the influence of full junctin on RyR1 and RyR2 activity. The C-terminal domain of junctin binds to residues including the S1-S2 linker of RyR1 and N-terminal domain of junctin binds between RyR1 residues 1078 and 2156.
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Linwei
authors.initials: L
authors.lastname: Li
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Shamaruh
authors.initials: S
authors.lastname: Mirza
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Spencer J
authors.initials: SJ
authors.lastname: Richardson
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Esther M
authors.initials: EM
authors.lastname: Gallant
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Chris
authors.initials: C
authors.lastname: Thekkedam
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Suzy M
authors.initials: SM
authors.lastname: Pace
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Francesco
authors.initials: F
authors.lastname: Zorzato
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia.
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Beard
authors.affiliation: John Curtin School of Medical Research, ACT 0200, Australia angela.dulhunty@anu.edu.au.
authors.forename: Angela F
authors.initials: AF
authors.lastname: Dulhunty
mesh_terms: D000818:Animals; D020013:Calcium Signaling / Q000502:physiology*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004285:Dogs; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics / Q000378:metabolism*
chemical_list: D002135:Calcium-Binding Proteins; D019837:Ryanodine Receptor Calcium Release Channel
keywords: ASPH; Cytoplasmic interaction; Junctin; Luminal interaction; RyR; Ryanodine receptor; Sarcoplasmic reticulum