Date of Award

12-2010

Document Type

Campus Access Thesis

Degree Name

Master of Science (MS)

Department

Biotechnology and Biomedical Science

First Advisor

Steven M. Ackerman

Second Advisor

Gregory Beck

Third Advisor

Manickam Sugumaran

Abstract

Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-ĸB), or Rel, proteins are important transcription factors critical to innate immune responses in many species, but this family of proteins has never been positively identified in plants. There is good reason, however, to speculate that transcription factors similar to NF-ĸB exist in plants, including significant identity between one plant protein (Nim1) and the inhibitor of NF-ĸB, IĸB. Our lab has been working on trying to isolate NF-ĸB-like transcription factors in plants. Assays such as Yeast One-Hybrid have proven unsuccessful in the past presumably because the transcription factors we are searching for are composed of heterodimers.

Analysis of the Arabidopsis thaliana genome reveals putative NF-ĸB DNA binding sites upstream of known genes At2g21850 and At2g40420. Our lab's latest efforts have focused on constructing radioactive probes containing these putative binding sites to ascertain whether a protein(s), presumably a transcription factor, will bind to these putative binding sites. Such a protein or protein complex could show identity to an NF-ĸB transcription factor or Nf-ĸB heterodimer.

A probe containing one of these putative NF-ĸB DNA binding sites upstream of gene At2g21850 binds to a complex of proteins found in nuclear protein extract purified from Triticum aestivum. This finding provides more evidence that NF-ĸB/Rel-like transcription factors may exist in plants.

In addition to the molecular methods utilized for this thesis work, the 5' region of the NIM1 protein, a protein which bears significant identity to an NF-ĸB inhibitor, was analyzed using a comparative genomics approach. The results of this analysis did not produce definitive results.

Comments

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