Abstract

The yeast ADA proteins and TBP-class of SPT proteins interact in multiple complexes (ADA/SPT complexes) that are required for the activation and repression of transcription. We have identified Tra1 as a component of these regulatory complexes through the tandem mass spectrometry analysis of proeints that associate with Ngg1/Ada3. Tra1 is a member of a group of putative protein kinases, that have acarboxyl-terminal region related to phosphatidylinisitol 3-kinases. The interaction between Tra1 and ADA/SPT components was verified by the reciprocal coimmiunprecipitation of HA-Ada2 and myc-Tra1 from whole cell extracts in one or complexes also containing Spt7. The association of Tra1 with more that one ADA/SPT complex was suggested by its cofractionation with Ada2, Ngg1 and Spt7 in two separate peaks on a FPLC mono Q column. In the absence of Ada2, the elution profile of Tra1p shifted to a single distinct peak. Tra1 also copurified with ADA/SPT proteins on a DNA-cellulose column where the binding of Tra1 to DNA-cellulose required the ADA components. As well as allowing the initial determination of Tra1p within the ADA/SPT complexes the tandem mass spectrometry analysis identified a phosphopeptide located near the protein's amino-terminus.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR011823-05
Application #
6348285
Study Section
Project Start
2000-09-01
Project End
2001-08-31
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
5
Fiscal Year
2000
Total Cost
$3,280
Indirect Cost

  Institution

Name
University of Washington
Department
Type
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Hollmann, Taylor; Kim, Tae Kwon; Tirloni, Lucas et al. (2018) Identification and characterization of proteins in the Amblyomma americanum tick cement cone. Int J Parasitol 48:211-224
Stieg, David C; Willis, Stephen D; Ganesan, Vidyaramanan et al. (2018) A complex molecular switch directs stress-induced cyclin C nuclear release through SCFGrr1-mediated degradation of Med13. Mol Biol Cell 29:363-375
Seixas, Adriana; Alzugaray, María Fernanda; Tirloni, Lucas et al. (2018) Expression profile of Rhipicephalus microplus vitellogenin receptor during oogenesis. Ticks Tick Borne Dis 9:72-81
Wang, Zheng; Wu, Catherine; Aslanian, Aaron et al. (2018) Defective RNA polymerase III is negatively regulated by the SUMO-Ubiquitin-Cdc48 pathway. Elife 7:
Xavier, Marina Amaral; Tirloni, Lucas; Pinto, Antônio F M et al. (2018) A proteomic insight into vitellogenesis during tick ovary maturation. Sci Rep 8:4698
Luhtala, Natalie; Aslanian, Aaron; Yates 3rd, John R et al. (2017) Secreted Glioblastoma Nanovesicles Contain Intracellular Signaling Proteins and Active Ras Incorporated in a Farnesylation-dependent Manner. J Biol Chem 292:611-628
Thakar, Sonal; Wang, Liqing; Yu, Ting et al. (2017) Evidence for opposing roles of Celsr3 and Vangl2 in glutamatergic synapse formation. Proc Natl Acad Sci U S A 114:E610-E618
Jin, Meiyan; Fuller, Gregory G; Han, Ting et al. (2017) Glycolytic Enzymes Coalesce in G Bodies under Hypoxic Stress. Cell Rep 20:895-908
Ogami, Koichi; Richard, Patricia; Chen, Yaqiong et al. (2017) An Mtr4/ZFC3H1 complex facilitates turnover of unstable nuclear RNAs to prevent their cytoplasmic transport and global translational repression. Genes Dev 31:1257-1271
Ju Lee, Hyun; Bartsch, Deniz; Xiao, Cally et al. (2017) A post-transcriptional program coordinated by CSDE1 prevents intrinsic neural differentiation of human embryonic stem cells. Nat Commun 8:1456

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