Three cDNAs that encode two-component phosphorelay-mediator-like proteins were cloned from Arabidopsis thaliana. Putative proteins (ATHP1-3) contain an HPt (Histidine-containing Phospho transfer)-like domain with a conserved histidine and some invariant residues that are involved in phosphorelay. Growth retardation of YPD1-disrupted yeast cells was reversed with ATHPs, which indicates that ATHPs function as phosphorelay mediators in yeast cells. The ATHP genes are expressed more in roots than in other tissues, similar to the expression of genes for a sensor histidine kinase, ATHK1, and response regulators ATRR1-4. These results suggest that ATHPs function as two-component phosphorelay mediators between sensor histidine kinase and response regulators in Arabidopsis.