Btn1 affects cytokinesis and cell-wall deposition by independent mechanisms, one of which is linked to dysregulation of vacuole pH.
btn1, the Schizosaccharomyces pombe orthologue of the human Batten-disease gene CLN3, is involved in vacuole pH homeostasis. We show that loss of btn1 also results in a defective cell wall marked by sensitivity to zymolyase, a beta-glucanase. The defect can be rescued by expression of Btn1p or CLN3, and the extent of the defect correlates with disease severity. The vacuole and cell-wall defects are linked by a common pH-dependent mechanism, because they are suppressed by growth in acidic pH and a similar glucan defect is also apparent in the V-type H(+) ATPase (v-ATPase) mutants vma1Delta and vma3Delta. Significantly, Btn1p acts as a multicopy suppressor of the cell-wall and other vacuole-related defects of these v-ATPase-null cells. In addition, Btn1p is required in a second, pH-independent, process that affects sites of polarised growth and of cell-wall deposition, particularly at the septum, causing cytokinesis problems under normal growth conditions and eventual cell lysis at 37 degrees C. Thus, Btn1p impacts two independent processes, which suggests that Batten disease is more than a pH-related lysosome disorder.
Codlin
Sandra
S
MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT.
Haines
Rebecca L
RL
Burden
J Jemima E
JJ
Mole
Sara E
SE
D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism*; D048749:Cytokinesis / Q000187:drug effects; D004705:Endocytosis / Q000187:drug effects; D017353:Gene Deletion; D016153:Genes, Suppressor; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D006867:Hydrolases / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism*; D009472:Neuronal Ceroid-Lipofuscinoses / Q000378:metabolism; D010641:Phenotype; D012568:Schizosaccharomyces / Q000166:cytology* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D013696:Temperature; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism; D014617:Vacuoles / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*
C509077:BTN1 protein, S pombe; D008565:Membrane Proteins; D029702:Schizosaccharomyces pombe Proteins; C037833:zymolyase; D006867:Hydrolases; D025262:Vacuolar Proton-Translocating ATPases