Centrosomal P4.1-associated Protein Is a New Member of Transcriptional Coactivators for Nuclear Factor-κB

Abstract

Nuclear factor-B (NF-B) is a transcription factor important for various cellular events such as inflammation, immune response, proliferation, and apoptosis. In this study, we performed a yeast two-hybrid screening using the N-terminal domain of the p65 subunit (RelA) of NF-B as bait and isolated centrosomal P4.1-associated protein (CPAP) as a candidate for a RelA-associating partner. Glutathione S-transferase pull-down assays and co-immunoprecipitation experiments followed by Western blotting also showed association of CPAP with RelA. When overexpressed, CPAP enhanced NF-B-dependent transcription induced by tumor necrosis factor-␣ (TNF␣). Reduction of the protein level of endogenous CPAP by RNA interference resulted in decreased activation of NF-B by TNF␣. After treatment with TNF␣, a portion of CPAP was observed to accumulate in the nucleus, although CPAP was found primarily in the cytoplasm without any stimulation. Moreover, CPAP was observed in a complex recruited to the transcriptional promoter region containing the NF-B-binding motif. One hybrid assay showed that CPAP has the potential to activate gene expression when tethered to the transcriptional promoter. These data suggest that CPAP functions as a coactivator of NF-B-mediated transcription. Since a physiological interaction between CPAP and the coactivator p300/CREB-binding protein was also observed and synergistic activation of NF-B-mediated transcription was achieved by these proteins, CPAPdependent transcriptional activation is likely to include p300/CREB-binding protein.

Cancer and embryo expression protein 65 promotes cancer cell growth and metastasis

“…CEP65 has been identified to contain eight coiled-coil domains (3), which are involved in the mediation of protein-protein interactions (32). To date, the spectrum of CEP65-binding proteins is unclear, while several other centrosomal proteins have been found to be involved in the regulation of gene transcription (5,33). For instance, NPHP6/CEP290 is physically associated with activating transcription factor 4 (ATF4), which may synergistically regulate renal development (5).…”

Interleukin-1 and Tumor Necrosis Factor-α Trigger Restriction of Hepatitis B Virus Infection via a Cytidine Deaminase Activation-induced Cytidine Deaminase (AID)

“…RT-PCR was performed as described previously (45) using a one-step RNA PCR kit (Takara). Primers for amplifying each gene were as follows: 5′-CTCTGAGGTTTAGCATTTCA-3′ and 5′-CTCCAGGTCCAAAATGAATA-3′ for cIAP ; 5′-GCAGATTTATCAACGGCTTT-3′ and 5′-CAGTTTTCCACCACAACAAA-3′ for XIAP ; 5′-TAGCCAACATGTCCTCACAGAC-3′ and 5′-TCTTCTACCACTGGTTTCATGC-3′ for ISG56 ; 5′-GCCTTTTCATCCAAATGGAATTC-3′ and 5′-GAAATCTGTTCTGGGCTCATG-3′ for PKR ; and 5′-CCATGGAGAAGGCTGGGG-3′ and 5′-CAAAGTTGTCATGGATGACC-3′ for GAPDH , respectively.…”

A novel deletion mutation in CENPJ gene in a Pakistani family with autosomal recessive primary microcephaly

“…The hydrophobic regions in CENPJ form a stripe that winds around alpha-helix, which is capable of interacting with the hydrophobic residues of a second molecule to form a coiled-coil structure (Cohen and Parry 1986;Hung et al 2000). Several recent reports have shown that CENPJ contributes in the RelA, STAT5 and NF-kB mediated transcription, but these seem less likely to influence neurogenesis (Peng et al 2002;Koyanagi et al 2005;Bond and Woods 2006).…”