Source: TEXAS A&M UNIVERSITY submitted to NRP
MOLECULAR MECHANISM OF MICROBIAL NUCLEIC ACIDS SENSING THROUGH STING
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0230777
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Aug 8, 2012
Project End Date
Aug 7, 2017
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001
Performing Department
Biochemistry & Biophysics
Non Technical Summary
In this research, we will identify proteins involved in antiviral immune responses. The functions of the new proteins will be studied by cell based assays and structural studies.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
72239991090100%
Knowledge Area
722 - Zoonotic Diseases and Parasites Affecting Humans;

Subject Of Investigation
3999 - Animal research, general;

Field Of Science
1090 - Immunology;
Goals / Objectives
The objective of this research is to identify new proteins involved in nucleic acids sensing and signaling via the adaptor protein STING and elucidate their roles in STING signaling at molecular level. This study will provide fundamental insights into the mechanism of microbial nucleic acid sensing by the immune system.
Project Methods
The techniques used in this research include: 1. Yeast two hybride screening of human cDNA library 2. Tandem Affinity purification of protein binding partners 3. Luciferase reporter assays in cells 4. RNA interference gene knockdown 4. X-ray crystallography of protein and protein complexes

Progress 08/08/12 to 08/07/17

Outputs
Target Audience:Scientists in the filed of molecular biology and immunology. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two graduate students and two post-docs received training in structural biology and molecular immunology. How have the results been disseminated to communities of interest?Yes, the papers were published in scientific journals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? This research elucidated the molecular mechanism of host defense by the cGAS-STING pathway.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Xin Guo, Chang Shu, Honggui Li, Ya Pei, Shih-Lung Woo, Juan Zheng, Mengyang Liu, Hang Xu, Rachel Botchlett, Ting Guo, Yuli Cai, Xinsheng Gao, Jing Zhou, Lu Chen, Qifu Li, Xiaoqiu Xiao, Linglin Xie, Ke K. Zhang, Jun-Yuan Ji, Yuqing Huo, Fanyin Meng, Gianfranco Alpini, Pingwei Li*, and Chaodong Wu* (2017). Cyclic GMP-AMP ameliorates diet-induced metabolic dysregulation and regulates proinflammatory responses distinctly from STING activation. Scientific Reports 7:6335, DOI: 10.1038/s41598-017-05884-y.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Zhonghui Zhang, Fuqu Hu, Min Woo Sung, Chang Shu, Claudia Castillo-Gonz�lez, Hisashi Koiwa, Guiliang Tang, Martin Dickman, Pingwei Li*, and Xiuren Zhang* (2017). RISC-Interacting Clearing 3- 5 Exoribonucleases (RICEs) degrade uridylated cleavage fragments to maintain functional RISC in Arabidopsis. eLife. doi:10.7554/eLife.24466
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Baoyu Zhao#, Guanghui Yi#, Fenglei Du#, Yin-Chih Chuang, Robert C. Vaughan, Banumathi Sankaran, C. Cheng Kao*, Pingwei Li* (2017). Structure and function of the Zika virus full-length NS5 protein. Nature Commun. 8, 14672, doi:10.1038/ncomms14762. (# These authors made equal contributions to this work; *Corresponding authors.)


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Scientists in the molecular biology of host defense by the immune system. Changes/Problems:We are developping techniques to affinity purify the STING containing signaling complex and plan to characterize that by mass spectrometry or structural approaches. What opportunities for training and professional development has the project provided?Two post-docs were trained in this research project. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?We are working on the moleclar mechanism of how the protein kinase TBK1 is recruited and activated by STING.

Impacts
What was accomplished under these goals? We have determined the structural basis of transcription factor IRF3 recruitment by STING using X-ray crystallography.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Baoyu Zhao, Chang Shu, Xinsheng Gao, Banumathi Sankaran, Fenglei Du, Catherine L. Shelton, Andrew B. Herr, Jun-Yuan Ji, and Pingwei Li* (2016). Structural basis for concerted recruitment and activation of IRF-3 by innate immune adaptor proteins. Proc Natl Acad Sci U S A 113, E3423-E3430. doi:10.1073/pnas.1603269113.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Our research is to elucidate the mechanism of how our body sense viral infection through the cGAS/STING pathway. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two post-docs were trained in this research. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We have recently elucidate the structural basis of interferon reglulatory factor IRF3 recruitment by phosphorylated STING. A manuscript on this was submitted recently.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Guanghui Yi, Yahong Wen, Chang Shu, Qinxian Han, Kouacou V. Konan, Pingwei Li, and C. Cheng Kao (2015). The Hepatitis C virus NS4B can suppress STING accumulation to evade innate immune responses. Journal of Virology, doi:10.1128/JVI.01720-15.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: We have been working on the structural studies of the mouse AIM2 dsDNA sensor for the last reporting year. We have determined the crystal structure of mouse AIM2 without bound DNA in two different crystal froms. We have als crystallized mouse AIM2 with a 12 nt single stranded DNA that may suppress the activation of AIM2 in vivo. Structural analysis of this complex is in progress. In addiiton, we have determined the crystal structure of a cytosolic DNA sensor cGAS that regulate the induction of type I interferons. Changes/Problems: Our future research will focus on the molecular and cellular mechanism of STING mediated signaling. What opportunities for training and professional development has the project provided? Two post-doc researchers were trained in our research in the field of structural biology. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? In the next stage of our research, we will investiage the roles of STING mediated signaling in antitumor immune responses.

Impacts
What was accomplished under these goals? We have determined the crystal structure of cGAS, a DNA sensor upstream of STING. In additon, we have detrmined the structure of STING bound to small molecule ligand DMXAA and CMX.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xin Li#, Chang Shu#, Guanghui Yi, Catherine T. Chaton, Catherine L. Shelton, Jiasheng Diao, Xiaobing Zuo, C Cheng Kao, Andrew B. Herr, and Pingwei Li* (2013). Cyclic GMP-AMP synthase is activated by double-stranded DNA-induced oligomerization. Immunity 39, 1019-103. (# These authors contributed equally to the work)


Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Basic life science research community. Changes/Problems: We have proposed to work on the DNA sensor AIM2. This project is largely done by us and others. Now we focus on another DNA sensor cGAS in innate immunity. What opportunities for training and professional development has the project provided? Three post-docs were trained in this project. How have the results been disseminated to communities of interest? The results were published in peer reviewed scientific journals What do you plan to do during the next reporting period to accomplish the goals? We will continue our research in the antimicrobial immunity

Impacts
What was accomplished under these goals? The major goal of this project is to elucidate the mechanism of innate immunity to dsDNA.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xin Li#, Chang Shu#, Guanghui Yi, Catherine T. Chaton, Catherine L. Shelton, Jiasheng Diao, Xiaobing Zuo, C Cheng Kao, Andrew B. Herr, and Pingwei Li* (2013). Cyclic GMP-AMP synthase is activated by double-stranded DNA-induced oligomerization. Immunity 39, 1019-103. (# These authors contributed equally to the work)
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Guanghui Yi, Volker P. Brendel, Chang shu, Pingwei Li, Satheesh Palanathan, and C. Cheng Kao (2013). Single nucleotide polymorphisms of human STING can affect innate immune response to cyclic dinucleotides. PLOS One, 8, e77846.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Chang Shu, Banumathi Sankaran, Catherine T. Chaton, Andrew B. Herr, Ashutosh Mishra, Junmin Peng, and Pingwei Li * (2013). Structural insights into the functions of TBK1 in innate antimicrobial immunity. Structure 21, 1137-1148.


Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: We have made very good progress in the research during the last half year. First, we have determined the structure of STING bound to a new ligand c-di-AMP. We have also crystallized the protein kinase TBK1 downstream of STING and determined the structure recently. The upstream sensor of STING was identified recently by others. We started structural studies of the DNA sensor and got crystals of the protein and collected diffractions data. We are also working on the mechanism of transcription activation by TBK1 in the signaling pathway. We have purified in vitro phosphorylated IRF3 and started biophysical characterization of the protein. PARTICIPANTS: Pingwei Li and two post-docs, Dr. Chang Shu and Dr. Xin Li. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
These studies will advance our understanding of how cells detect DNA from the pathogen and have potential applications in drug design.

Publications

  • Cheng Lu, Pingwei Li (2012). Preparation of short RNA by in vitro transcription. Methods Mol. Biol, Edited by Graeme L. Conn, Humana Press, 941, 59-68.
  • Min Woo Sung, Tylan Watts, Pingwei Li* (2012). Crystallographic characterization of mouse AIM2-HIN200 domain bound to a 15 bp and an 18 bp double-stranded DNA. Acta Crystallogr Sect F Struct Biol Cryst Commun. F68, 1081-1084.
  • Chang Shu, Guanghui Yi, Tylan Watts, C. Cheng Kao, Pingwei Li* (2012). Structure of STING bound to c-di-GMP reveals the mechanism of cyclic dinucleotide recognition by the immune system. Nature Structural and Molecular Biology 19, 722-724.