Membrane-Fluidization-Dependent and -Independent Pathways Are Involved in Heat-Stress-Inducible Gene Expression in the Marine Red Alga Neopyropia yezoensis

Heat stress responses are complex regulatory processes, including sensing, signal transduction, and gene expression. However, the exact mechanisms of these processes in seaweeds are not well known. We explored the relationship between membrane physical states and gene expression in the red alga Neopyropia yezoensis. To analyze heat-stress-induced gene expression, we identified two homologs of the heat-inducible high temperature response 2 (HTR2) gene in Neopyropia seriata, named NyHTR2 and NyHTR2L. We found conservation of HTR2 homologs only within the order Bangiales; their products contained a novel conserved cysteine repeat which we designated the Bangiales cysteine-rich motif. A quantitative mRNA analysis showed that expression of NyHTR2 and NyHTR2L was induced by heat stress. However, the membrane fluidizer benzyl alcohol (BA) did not induce expression of these genes, indicating that the effect of heat was not due to membrane fluidization. In contrast, expression of genes encoding multiprotein-bridging factor 1 (NyMBF1) and HSP70s (NyHSP70-1 and NyHSP70-2) was induced by heat stress and by BA, indicating that it involved a membrane-fluidization-dependent pathway. In addition, dark treatment under heat stress promoted expression of NyHTR2, NyHTR2L, NyMBF1, and NyHSP70-2, but not NyHSP70-1; expression of NyHTR2 and NyHTR2L was membrane-fluidization-independent, and that of other genes was membrane-fluidization-dependent. These findings indicate that the heat stress response in N. yezoensis involves membrane-fluidization-dependent and -independent pathways.