TMPIT1 from wild emmer wheat: First characterisation of a stress-inducible integral membrane protein

In this study a gene for a drought stress-inducible putative membrane protein was cloned and characterised from root tissue of wild emmer wheat. Sequence analysis indicated that the protein is a member of the widespread but hitherto uncharacterised TMPIT (transmembrane protein inducible by TNF-α) family, so it was labelled TdicTMPIT1. Real-time RT-PCR showed that the TdicTMPIT1 gene is upregulated on drought stress in drought-tolerant wild emmer wheat, but not in a drought-sensitive accession or in cultivated durum wheat. The TdicTMPIT1 product was predicted to be a membrane protein with four transmembrane helices. The protein was expressed and analysed in Escherichia coli and Saccharomyces cerevisiae. Cellular localisation of the protein in the cell was also investigated using an eGFP-tagged form of the protein in S. cerevisiae. Results obtained by confocal laser microscopy indicated that the TdicTMPIT1 tagged with GFP was localised in a membraneous compartment. It is concluded that TdicTMPIT1 is a membrane protein associated with the drought stress response in wild emmer wheat, and so it may be useful for the improvement of modern wheat genotypes. Members of this protein family in other organisms are proposed also to be involved in stress responses.