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1 December 2015 N- and C-terminal non-conserved residues contribute to transactivation by a sea anemone ( Nematostella vectensis) NF-κB transcription factor
Anar Alshanbayeva, Ajith Thomas, Martine Tremblay, Majed Abbas, Fatema Abdurrob, Talal Almojel, Alexis Aparicio, Dhishant Asarpota, Ashley Ayers, Amar Aziz, Jacob Bishop, Theresa Christie, Ming Jie, Melissa Chua, Oliver Chung, Nikhil Dhar, Alexa Diedrich, Chelsea Fortin, Qianjing He, Sarah Heerboth, Rosanna Hok, Aditya Khedkar, Shweta Kitchloo, Caitlin Lawlor, Brian Leonard, Shannon Linderman, Mariam Maloyan, Lauren Miller, Christine Pak, Abishy Pandita, Isabel Park, Neil Patel, Janani Ramachandran, Marinaliz Reynoso, Yasmina Samaha, Grace Thole, Jason Turnbill, Linge Xia, Jingzhi Zhu, Caryn Navarro, Thomas D. Gilmore
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Abstract

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.

Anar Alshanbayeva, Ajith Thomas, Martine Tremblay, Majed Abbas, Fatema Abdurrob, Talal Almojel, Alexis Aparicio, Dhishant Asarpota, Ashley Ayers, Amar Aziz, Jacob Bishop, Theresa Christie, Ming Jie, Melissa Chua, Oliver Chung, Nikhil Dhar, Alexa Diedrich, Chelsea Fortin, Qianjing He, Sarah Heerboth, Rosanna Hok, Aditya Khedkar, Shweta Kitchloo, Caitlin Lawlor, Brian Leonard, Shannon Linderman, Mariam Maloyan, Lauren Miller, Christine Pak, Abishy Pandita, Isabel Park, Neil Patel, Janani Ramachandran, Marinaliz Reynoso, Yasmina Samaha, Grace Thole, Jason Turnbill, Linge Xia, Jingzhi Zhu, Caryn Navarro, and Thomas D. Gilmore "N- and C-terminal non-conserved residues contribute to transactivation by a sea anemone ( Nematostella vectensis) NF-κB transcription factor," BIOS 86(4), 165-175, (1 December 2015). https://doi.org/10.1893/0005-3155-86.4.165
Received: 4 August 2014; Accepted: 1 November 2014; Published: 1 December 2015
JOURNAL ARTICLE
11 PAGES


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NF-kappaB
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