Human herpesvirus 6 (HHV-6) immediate-early (IE) 2 protein (IE2) may play important but incompletely defined roles during infection. We used yeast two-hybrid screening to detect proteins interacting with HHV-6 IE2, and found heterogeneous nuclear ribonucleoprotein K (hnRNP K) and the beta subunit of casein kinase 2 (CK2beta) specifically interacted with HHV-6 IE2. The interactions were confirmed by GST pull-down assay, coimmunoprecipitation, and colocalization studies. These findings indicate that the HHV-6 IE2 protein interacts with hnRNP K and CK2, and these interactions may affect viral and cellular RNA transcription and translation in viral replication.