Very little is known about SMYD2 (SET and MYND containing protein 2), a member of the SMYD protein family. However, the interest in better understanding the roles of SMYD2 has grown due to recent reports indicating that SMYD2 methylates p53 and histone H3. In this paper, we present a combined proteomic and genomic study of SMYD2 designed to elucidate its molecular roles. We report the cytosolic and nuclear interactome of SMYD2 using a combination of IP-HTMS, ChIP-HTMS, and co-immunoprecipitation methods. In particular, we report that SMYD2 interacts with HSP90 independently of the SET and MYND domain; with EBP41L3 through the MYND domain; and with p53 through the SET domain. We demonstrated that the interaction of SMYD2 with HSP90 enhances SMYD2 histone methyltransferase activity and specificity for histone H3 at lysine 4 (H3K4) in vitro. Interestingly, histone H3K36 methyltransferase activity is independent of its interaction with HSP90 similarly to LSD1 dependency on the androgen receptor. We also show that the SET domain is required for the methylation at H3K4. We demonstrated using a modified chromatin IP protocol that the SMYD2 gain of function leads to an increase in H3K4 methylation in vivo, while no observable levels of H3K36 were observed. We also report that the SMYD2 gain of function is correlated with the up-regulation of 37 and down-regulation of 4 genes, the majority of which are involved in the cell cycle, chromatin remodeling, and transcriptional regulation. We validate that the up-regulation of TACC2 by SMYD2 occurs as a result of SMYD2 binding to the TACC2 promoter where it methylates H3K4. Furthermore, the combination of the SMYD2 interactome with the gene expressions data suggests that some of the genes regulated by SMYD2 are closely associated with SMYD2 interacting proteins.