The 5' untranslated region of potato SBgLR gene contributes to pollen-specific expression.

ABSTRACT

MAIN CONCLUSION: The 5'UTR of SBgLR enhances gene expression by regulating both its transcription and translation. SBgLR (Solanum tuberosum genomic lysine rich) is a pollen-specific gene in Solanum tuberosum that encodes a microtubule-associated protein. The region from -85 to +180 (transcription start site at +1) was determined to be critical for specific expression in pollen grains. Transient and stable expression assays showed that the 5'UTR (from +1 to +184) enhanced gene expression in all detected tissues of transgenic tobacco. Deletion analysis demonstrated that the secondary structure of the 5'UTR had no effect on pollen-specific SBgLR expression, while the region from +31 to +60 was crucial. Further investigation indicated that mRNA expression was slightly decreased when the +31 to +60 region was deleted, but the mRNA decay rate remained unchanged. Mutation analysis also confirmed that the pollen-specific element TTTCT, located at +37, played an important role in pollen-specific expression. Using yeast one-hybrid screening, we isolated a DNA-binding with one finger (Dof) protein gene (StDof23) and an AT-hook motif nuclear-localized (AHL) protein gene (StAHL) from potato pollen. Further investigation indicated that StDof23 interacted with and positively regulated the +31 to +60 region; moreover, StAHL interacted with and negatively regulated the -49 to +60 region. These results demonstrate that the 5'UTR not only enhanced gene expression but also altered the tissue-specific expression pattern by regulating both transcription and translation.