Yeast Two-Hybrid Screens to Identify Drosophila PP1-Binding Proteins

2007 ◽  
pp. 155-180 ◽  
Author(s):  
Daimark Bennett ◽  
Luke Alphey
Keyword(s):  
Binding Proteins ◽  
Yeast Two Hybrid ◽  
Two Hybrid

Identification of Arabidopsis CCR4-NOT Complexes with Pumilio RNA-Binding Proteins, APUM5 and APUM2

2019 ◽  
Vol 60 (9) ◽  
pp. 2015-2025 ◽  
Author(s):  
Toshihiro Arae ◽  
Kotone Morita ◽  
Riko Imahori ◽  
Yuya Suzuki ◽  
Shigetaka Yasuda ◽  
...  
Keyword(s):  
Gene Expression ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Physiological Effect ◽  
Yeast Two Hybrid ◽  
Recognition Mechanism ◽  
Gene Expression Control ◽  
Core Components ◽  
Two Hybrid

Abstract CCR4/CAF1 are widely conserved deadenylases in eukaryotes. They form a large complex that includes NOT1 as a scaffold protein and various NOT proteins that are core components of multiple levels of gene expression control. The CCR4-NOT complex also contains several RNA-binding proteins as accessory proteins, which are required for target recognition by CCR4/CAF1 deadenylases. AtCCR4a/b, orthologs of human CCR4 in Arabidopsis, have various physiological effects. AtCCR4 isoforms are likely to have specific target mRNAs related to each physiological effect; however, AtCCR4 does not have RNA-binding capability. Therefore, identifying factors that interact with AtCCR4a/b is indispensable to understand its function as a regulator of gene expression, as well as the target mRNA recognition mechanism. Here, we identified putative components of the AtCCR4-NOT complex using co-immunoprecipitation in combination with mass spectrometry using FLAG-tagged AtCCR4b and subsequent verification with a yeast two-hybrid assay. Interestingly, four of 11 AtCAF1 isoforms interacted with both AtCCR4b and AtNOT1, whereas two isoforms interacted only with AtNOT1 in yeast two-hybrid assays. These results imply that Arabidopsis has multiple CCR4-NOT complexes with various combinations of deadenylases. We also revealed that the RNA-binding protein Arabidopsis Pumilio 5 and 2 interacted with AtCCR4a/b in the cytoplasm with a few foci.

scholarly journals Screening of TMEM48 binding proteins in testis by yeast two-hybrid method

2013 ◽  
Vol 41 (2) ◽  
pp. 77-86
Author(s):  
Shimpei KAJITA ◽  
Kouyou AKIYAMA ◽  
Takehito TSUJI ◽  
Tetsuo KUNIEDA
Keyword(s):  
Hybrid Method ◽  
Binding Proteins ◽  
Yeast Two Hybrid ◽  
Two Hybrid

scholarly journals Yeast two-hybrid analysis of the origin recognition complex ofSaccharomyces cerevisiae: interaction between subunits and identification of binding proteins

2007 ◽  
Vol 7 (8) ◽  
pp. 1263-1269 ◽  
Author(s):  
Kazuya Matsuda ◽  
Masaki Makise ◽  
Yoshihiro Sueyasu ◽  
Masaya Takehara ◽  
Teita Asano ◽  
...  
Keyword(s):  
Origin Recognition Complex ◽  
Binding Proteins ◽  
Yeast Two Hybrid ◽  
Hybrid Analysis ◽  
Two Hybrid ◽  
Origin Recognition

scholarly journals The Mdm2 Oncoprotein Interacts with the Cell Fate Regulator Numb

1998 ◽  
Vol 18 (7) ◽  
pp. 3974-3982 ◽  
Author(s):  
Tamar Juven-Gershon ◽  
Ohad Shifman ◽  
Tamar Unger ◽  
Adi Elkeles ◽  
Ygal Haupt ◽  
...  
Keyword(s):  
Tumor Suppressor ◽  
Tumor Cells ◽  
Cell Fate ◽  
Binding Proteins ◽  
Yeast Two Hybrid ◽  
Transfected Cells ◽  
P53 Tumor Suppressor ◽  
Two Hybrid

ABSTRACT The Mdm2 oncoprotein is a well-known inhibitor of the p53 tumor suppressor, but it may also possess p53-independent activities. In search of such p53-independent activities, the yeast two-hybrid screen was employed to identify Mdm2-binding proteins. We report that in vitro and in transfected cells, Mdm2 can associate with Numb, a protein involved in the determination of cell fate. This association causes translocation of overexpressed Numb into the nucleus and leads to a reduction in overall cellular Numb levels. Through its interaction with Numb, Mdm2 may influence processes such as differentiation and survival. This could potentially contribute to the altered properties of tumor cells which overexpress Mdm2.

scholarly journals A Yeast Heterogeneous Nuclear Ribonucleoprotein Complex Associated With RNA Polymerase II

Genetics ◽  
2000 ◽  
Vol 154 (2) ◽  
pp. 557-571 ◽  
Author(s):  
Nicholas K Conrad ◽  
Scott M Wilson ◽  
Eric J Steinmetz ◽  
Meera Patturajan ◽  
David A Brow ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Temperature Sensitive ◽  
Yeast Two Hybrid ◽  
Pol Ii ◽  
Physical Interactions ◽  
Two Hybrid

Abstract Recent evidence suggests a role for the carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II (pol II) in pre-mRNA processing. The yeast NRD1 gene encodes an essential RNA-binding protein that shares homology with mammalian CTD-binding proteins and is thought to regulate mRNA abundance by binding to a specific cis-acting element. The present work demonstrates genetic and physical interactions among Nrd1p, the pol II CTD, Nab3p, and the CTD kinase CTDK-I. Previous studies have shown that Nrd1p associates with the CTD of pol II in yeast two-hybrid assays via its CTD-interaction domain (CID). We show that nrd1 temperature-sensitive alleles are synthetically lethal with truncation of the CTD to 9 or 10 repeats. Nab3p, a yeast hnRNP, is a high-copy suppressor of some nrd1 temperature-sensitive alleles, interacts with Nrd1p in a yeast two-hybrid assay, and coimmunoprecipitates with Nrd1p. Temperature-sensitive alleles of NAB3 are suppressed by deletion of CTK1, a kinase that has been shown to phosphorylate the CTD and increase elongation efficiency in vitro. This set of genetic and physical interactions suggests a role for yeast RNA-binding proteins in transcriptional regulation.

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