Alternative splicing of an exitron determines the subnuclear localization of the Arabidopsis DNA‐glycosylase MBD4L under heat stress (SEGUNDA AUTORIA COMPARTIDA)

CECCHINI, NICOLÁS MIGUEL; TORRES, JOSÉ ROBERTO; LESCANO LÓPEZ, IGNACIO; COBO, SANTIAGO; NOTA, FLORENCIA; ALVAREZ, MARÍA ELENA

PLANT JOURNAL

WILEY-BLACKWELL PUBLISHING, INC

Año: 2022

0960-7412

DNA glycosylases are critical enzymes that recognize small base lesions in DNA and trigger their repair to preserve genome integrity. The Arabidopsis MBD4-like (MBD4L) DNA glycosylase improves tolerance to genotoxic stress. This enzyme is encoded by a single gene carrying an exitron at its 5´ region. Although alternative splicing (AS) of exitrons (protein-coding cryptic introns within exons) is suspected to increase protein diversity, phenotypes associated to exitron removal or retention are only known for a few genes. Here, we show that alternative splicing of the MBD4L exitron determines the generation of two enzyme isoforms with different subnuclear localization. Both isoforms conserve the catalytic domain but are directed either to the nucleoplasm or the nucleolus. Interestingly, heat-stress regulates the AS of the MBD4L exitron and increases the relative abundance of the nucleolar variant. This process depends on the splicing factors NTR1 and RS31. Notoriously, we generated transgenic plants expressing a mutated MBD4L-GFP gene version that abolished exitron splicing and found that nucleolar protein targeting was impaired in these plants. Our findings suggest that AS of the MBD4L exitron can function as a mechanism to drive the enzyme to the nucleolus during heat stress. Several DNA repair enzymes reach the nucleolus under particular conditions although AS of exitrons has not been so far associated with this process. To our knowledge, this is the first example of an exitron mediating enzyme localization in eukaryotes.