Journal Article

N- and C-terminal non-conserved residues contribute to transactivation by a sea anemone (Nematostella vectensis) NF-κB transcription factor

Turnbill, Jason, Samaha, Yasmina, Maloyan, Mariam, Dhar, Nikhil, Reynoso, Marinaliz, Chung, Oliver, Pandita, Abishy, Aparicio, Alexis, Ayers, Ashley, Hok, Rosanna, Park, Isabel, Bishop, Jacob, Patel, Neil, Navarro, Caryn, Thole, Grace, Xia, Linge, Chua, Ming Jie, Melissa, Heerboth, Sarah, Fortin, Chelsea, Khedkar, Aditya, Alshanbayeva, Anar, Zhu, Jingzhi, Pak, Christine, Kitchloo, Shweta, Leonard, Brian, Ramachandran, Janani, Gilmore, Thomas D., Christie, Theresa, Almojel, Talal, Miller, Lauren, Lawlor, Caitlin, Aziz, Amar, Asarpota, Dhishant, Tremblay, Martine, Thomas, Ajith, Linderman, Shannon, Diedrich, Alexa, Abbas, Majed, Abdurrob, Fatema, He, Qianjing

2015

Overview

NF-κB is an evolutionarily conserved eukaryotic transcription factor that plays a role in many important developmental and immune-related processes by activating target gene expression. The goal of these experiments was to define the sequences required for a sea anemone NF-κB's intrinsic transactivation activity by using mutant proteins with serial deletions of the N- and C-terminal sequences. Deletion mutants were constructed that were missing the C-terminal 15, 32 or 47 amino acids (aa) or the N-terminal 17, 27 or 47 aa of the 440 aa NF-κB protein from the starlet sea anemone, Nematostella vectensis (Nv), a simple model organism in the phylum Cnidaria. These Nv-NF-κB mutants were expressed as GAL4 fusion proteins in yeast, and their transactivation activities were assessed by LacZ reporter gene assays. The deletion of 47 aa from either the N terminus or the C terminus of NF-κB completely inactivates the transactivation function of Nv-NF-κB. In addition, we identified proline-258 in the center of the protein as a key residue for the transactivation function of Nv-NF-κB. Taken together, these results demonstrate that non-conserved N- and C-terminal residues are both required for the transcriptional activating function of the sea anemone NF-κB protein, suggesting that it has a novel functional domain structure among known NF-κB proteins.

• Publisher: Beta Beta Beta Biological Society
• Subject: amino acids / Anthozoa / Codons / DNA / gene expression / Genetic mutation / Journalism / mutants / NF-kappaB / Plasmids / Proteins / Reporter genes / Research Article / Transactivation / transcription (genetics) / transcription factor NF-kappa B / Transcription factors / transcriptional activation / Yeasts