Abstract: We used sequence motifs conserved in S-adenosylmethionine-dependent methyltransferases to identify 26 putative methyltransferases from the complete genome of the yeast Saccharomyces cerevisiae. Seven sequences with the best matches to the methyltransferase consensus motifs were selected for further study. We prepared yeast disruption mutants of each of the genes encoding these sequences, and we found that disruption of the YJL125c gene is lethal, whereas disruptions of YCR047c and YDR140w lead to slow growth phenotypes. Normal growth was observed when the YDL201w, YDR465c, YHR209w, and YOR240w genes were disrupted. Initial analysis of protein methylation patterns of all mutants by amino acid analysis revealed that the YDR465c mutant has a defect in the methylation of the delta-nitrogen atom of arginine residues. We propose that YDR465c codes for the methyltransferase responsible for this recently characterized type of protein methylation, and we designate the enzyme as Rmt2 (protein arginine methyltransferase). In addition, we show that the methylation of susceptible residues in Rmt2 substrates is likely to take place on nascent polypeptide chains and that these substrates exist in the cell as fully methylated species. Interestingly, Rmt2 has 27% sequence identity over 138 amino acids to the mammalian guanidinoacetate N-methyltransferase, an enzyme responsible for methylating the delta-nitrogen of the small molecule guanidinoacetate.