A novel and stress adaptive alternative oxidase derived from alternative splicing of duplicated exon in oyster Crassostrea virginica

Alternative oxidase (AOX) is a mitochondrial inner-membrane oxidase that accepts electrons directly from ubiquinol and reduces oxygen to water without involving cytochrome-linked electron transport chain. It is highly conserved in many non-vertebrate taxa and may protect cells against hypoxia and oxidative stress. We identified two AOX mRNAs in eastern oyster Crassostrea virginica, CvAOXA and CvAOXB, which differ by 170 bp but encode AOXs of the same size. Sequence analyses indicate that CvAOX has 10 exons with a tandem duplication of exon 10, and 3′ alternative splicing using either the first or second exon 10 produces the two variants CvAOXB or CvAOXA, respectively. The second exon 10 in CvAOXA is more conserved across taxa, while the first exon 10 in CvAOXB contains novel mutations surrounding key functional sites. Both variants are expressed in all organs with the expression of CvAOXA higher than that of CvAOXB under normal condition. Under stress by air exposure, CvAOXB showed significantly higher expression than CvAOXA and became the dominant variant. This is the first case of alternative splicing of duplicated exon in a mollusc that produces a novel variant adaptive to stress, highlighting genome’s versatility in generating diversity and phenotypic plasticity.