Enhanced expression of valosin-containing protein (VCP) correlates with invasiveness and metastatic potential of cancer cells; therefore its transcriptional regulation could be a new target for cancer therapy. Here, the mechanism for transactivation of VCP promoter by E74-like factor 2 (ELF2), a transcription factor which enhances VCP transcription, was investigated. There was a highly conserved region among species in VCP promoter with one proven motif for ELF2 binding. However, the mutation at this motif did not obviously change the promoter activity. Further survey revealed another ELF2 binding motif in the upstream of the conserved region. The promoter activity was modestly reduced when this upstream motif was mutated, and completely abolished when both upstream and downstream motifs were mutated. Chromatin immunoprecipitation assay revealed that ELF2 bound both upstream and downstream motifs. These findings indicate that ELF2 transactivates VCP promoter through binding to two motifs, with a predominant contribution of the upstream one.