Progress 09/01/03 to 08/30/09
Outputs
OUTPUTS: Activities: Conducting and analyzing experiments in drought tolerance Events: Have given local UNL seminars Services: technical consulting Products: Bought Illumina GA II sequencing system for general use as a core facility for both company and academic use. One graduate student complete MS thesis. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
New insights into plant cell signaling generated through protein kinase protein interaction studies. New insights into how chromatin modifications change during dehydration stress and how cell signaling involves chromatin changes.
Publications
- Ding X., Richter T., Chen M., Fujii H., Su Seo Y., Xie M., Zheng X., Kanrar S., Stevenson R. A., Dardick C., Li Y., Jiang H., Zhang Y., Yu F., Bartley L. E., Chern M., Bart R., Chen X., Zhu L., Farmerie W. G., Gribskov M., Zhu J-K., Fromm M.E., Ronald P. C., and Song W-Y. A Rice Kinase-Protein Interaction Map. Plant Physiol. 2009 Mar;149(3):1478-92.
- Ding Y, Lapko H, Ndamukong I, Xia Y, Al-Abdallat A, Lalithambika S, Sadder M, Saleh A, Fromm M, Riethoven J-J, Lu G, Avramova Z (2009) The Arabidopsis chromatin modifier ATX1, the myotubularin-like AtMTM, and the response to drought. Plant Signaling & Behavior. 4:1-10
- Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald L. Cerny, Chris Dardick, Patrick Canlas, Hiroaki Fujii, Michael Gribskov, Siddhartha Kanrar, Lucas Knoflicek, Becky Stevenson, Mingtang Xie, Xia Xu, Xianwu Zheng, Jian-Kang Zhu, Pamela Ronald, and Michael E. Fromm . Protein-Protein Interactions of Tandem Affinity Purified Protein Kinases from Rice. (2009 accepted PLoS ONE).
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Progress 10/01/07 to 09/30/08
Outputs
OUTPUTS: Lectures at UNL on the impact of biotechnology on agriculture. Co-teacher on new course on using new high throughput DNA sequencing for analyzying complex biological processes. Consultant to local industry on using genomics methods. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Provide basic knowledge to researchers in plant stress to help advance plant breeding and genetic engineering for improved drought tolerance. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Progress in protein kinase signaling in cereals has been achieved. Progress in new whole genome methods for analyzing chromatin structure during drought stress has been achieved. Implementation of new high throughput genomics methods locally.
Publications
- No publications reported this period
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Progress 10/01/06 to 09/30/07
Outputs
OUTPUTS: Protein interactions provided at website: http://rkd.ucdavis.edu/ which is a rice protein kinase database.
PARTICIPANTS: Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald Cerny, Chris Dardick, Patrick Canlas, Xia Xu, Michael Gribskov, Siddhartha Kanrar, Jian-Kang Zhu, Pamela Ronald, and Michael E. Fromm
TARGET AUDIENCES: Scientists in plant science in response to environmental stress
Impacts
Increased knowledge of protein kinase protein interactions to better understand signaling mechanisms in plants
Publications
- Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald Cerny, Chris Dardick, Patrick Canlas, Xia Xu, Michael Gribskov, Siddhartha Kanrar, Jian-Kang Zhu, Pamela Ronald, and Michael E. Fromm. Mass spectrometry identification of protein complexes from the purification of TAP-tagged protein kinases from transgenic rice plants. Plant J. 2006 Apr;46(1):1-13.
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Progress 10/01/05 to 09/30/06
Outputs
Our project to find proteins that interact with rice protein kinases is continuing. Forty-one rice protein kinase cDNAs encoding protein kinases were fused to the Tandem Affinity Purification (TAP) tag and expressed in transgenic rice plants. The TAP-tagged kinases and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by mass spectrometry (MS). Ninety-five percent of the TAP-tagged kinases were recovered. Fifty-six percent of the TAP-tagged kinases were found to interact with other rice proteins. A number of these interactions were consistent with known protein complexes found in other species, validating the TAP-tag method in rice plants. Phosphorylation sites were identified on four of the kinases that interacted with either 14-3-3 proteins or cyclins.
Impacts
Improved breeding and genetic engineering of cereals will require more detailed knowledge of the function of proteins and the mechansims of how they work. Determining protein-protein interactions is an important step in understanding a protein's function in the cell. Environmental and disease stresses are the major limiting factors in plant production and gaining an understanding of how plants respond to these stresses is key to improving their yields under stress.
Publications
- Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald Cerny, Chris Dardick, Patrick Canlas, Xia Xu, Michael Gribskov, Siddhartha Kanrar, Jian-Kang Zhu, Pamela Ronald, and Michael E. Fromm. 2006. Mass spectrometry identification of protein complexes from the purification of TAP-tagged protein kinases from transgenic rice plants. Plant J. in press.
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Progress 10/01/03 to 09/30/04
Outputs
The protein-protein interactions of rice protein kinases is being investigated. The goal is to gain a better understanding of plant cell signaling in cereals, with the long term goal of improving the cereals stress tolerance to environmental and disease stresses. over 100 rice protein kinases have been cloned, modified and screened in either rice plants or yeast two hybrids screens. These results will be published and posted on the website.
Impacts
Improved breeding and genetic engineering of cereals will require more detailed knowledge of the function of proteins and the mechansims of how they work. Determining protein-protein interactions is an important step in understanding a protein's function in the cell. Environmental and disease stresses are the major limiting factors in plant production and gaining an understanding of how plants respond to these stresses is key to improving their yields under stress.
Publications
- Rohila JS, Chen M, Cerny R, Fromm ME. (2004 ) Improved tandem affinity purification tag and methods for isolation of protein heterocomplexes from plants. Plant J. 38(1):172-81.
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