Functional dissection of the ADR1 CCR domain and identification of its protein interactions in the plant immune system
R-NB-LRR proteins are able to activate resistance signaling, characterized by hypersensitive response (HR). With little known of this domain, testing is required to find proteins that interact with this domain as well as its functional regions. To discover the functional regions, different six amino acid regions were replaced by a NAAIRS sequence through PCR mutagenesis and infiltrated into Nicotiana tabacum and N. benthamiana to determine how it affects HR. To find interactions, the R domain was to bind to potential protein interactors to induce transcription of a reporter gene. Of 18 mutations 13 were found to completely eradicate HR and the remaining found to display some degree of HR. From the Y2H assay, 31 possible protein interactors were found to interact with the R domain. The mutants revealed the different functional regions of this domain and these HR-abolishing mutations may abolish interactions with the true interactors, which will open up a stronger picture of how these pathways work.
Overall this experience was very eye-opening. It allowed me to finally enter the realm of research and working as a contributor to a larger project at hand, while still keeping control of your smaller project. I was able to peek into the world of what research might be like in the future and was able to work alongside people who understood the topic at hand in depth. Alongside working with your project the variety of speakers and seminars also give you a wide breadth of insight into a variety of topics in plant genetics. With all of these, the fellow interns who follow right alongside you, create a very welcoming experience where you are surrounded by many like-minded and motivated people. This research-filled summer has definitely become a lasting experience.