JAB1/CSN5 interacts with the GAL4 DNA binding domain: A note of caution about two-hybrid interactions

Nordgård, Oddmund; Dahle, Øyvind; Andersen, Tor Øyvind; Gabrielsen, Odd S.

doi:10.1016/s0300-9084(01)01329-3

Cited by 29 publications

(20 citation statements)

References 28 publications

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“…It has been previously reported that CSN5 interacts with the GAL4 DNA binding domain (Nordgå rd et al, 2001); thus, the isolation of CSN5 in a GAL4-based yeast two-hybrid screening should be considered cautiously. However, CSN5 homologs from different organisms have been isolated in this kind of screening in several independent studies (Kameda et al, 2006;Cho et al, 2008;Tanguy et al, 2008), and the interactions have been confirmed by other methods.…”

Section: C2 Interacts With the Plant Csn5mentioning

confidence: 99%

Geminiviruses Subvert Ubiquitination by Altering CSN-Mediated Derubylation of SCF E3 Ligase Complexes and Inhibit Jasmonate Signaling inArabidopsis thaliana

et al. 2011

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Viruses must create a suitable cell environment and elude defense mechanisms, which likely involves interactions with host proteins and subsequent interference with or usurpation of cellular machinery. Here, we describe a novel strategy used by plant DNA viruses (Geminiviruses) to redirect ubiquitination by interfering with the activity of the CSN (COP9 signalosome) complex. We show that geminiviral C2 protein interacts with CSN5, and its expression in transgenic plants compromises CSN activity on CUL1. Several responses regulated by the CUL1-based SCF ubiquitin E3 ligases (including responses to jasmonates, auxins, gibberellins, ethylene, and abscisic acid) are altered in these plants. Impairment of SCF function is confirmed by stabilization of yellow fluorescent protein-GAI, a substrate of the SCF SLY1 . Transcriptomic analysis of these transgenic plants highlights the response to jasmonates as the main SCF-dependent process affected by C2. Exogenous jasmonate treatment of Arabidopsis thaliana plants disrupts geminivirus infection, suggesting that the suppression of the jasmonate response might be crucial for infection. Our findings suggest that C2 affects the activity of SCFs, most likely through interference with the CSN. As SCFs are key regulators of many cellular processes, the capability of viruses to selectively interfere with or hijack the activity of these complexes might define a novel and powerful strategy in viral infections.

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Section: C2 Interacts With the Plant Csn5mentioning

confidence: 99%

Geminiviruses Subvert Ubiquitination by Altering CSN-Mediated Derubylation of SCF E3 Ligase Complexes and Inhibit Jasmonate Signaling inArabidopsis thaliana

et al. 2011

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“…COP9 subunit 5 and JAB1 are closely related; both have been found as the result of two-hybrid screens with several different bait proteins (Tomoda et al, 1999;Bianchi et al, 2000;Chauchereau et al, 2000;Li et al, 2000;Schwechheimer et al, 2001). There have even been suggestions that CSN-5 binds nonspecifically in the GAL4 system (Nordgard et al, 2001). In part to address that concern, we produced GST-CSN-5 and were able to successfully pull-down GLH-1 from worm homogenates, using infected GST alone and uninfected cell lysates as negative controls (Fig.…”

Section: Csn-5: a Modulator Of Several Proteins In The C Elegans Germentioning

confidence: 99%

The GLH Proteins, Caenorhabditis elegans P Granule Components, Associate with CSN-5 and KGB-1, Proteins Necessary for Fertility, and with ZYX-1, a Predicted Cytoskeletal Protein

Smith

Leung-Chiu

Montgomery

et al. 2002

Developmental Biology

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The GLH proteins belong to a family of four germline RNA helicases in Caenorhabditis elegans. These putative ATP-dependent enzymes localize to the P granules, which are nonmembranous complexes of protein and RNA exclusively found in the cytoplasm of all C. elegans germ cells and germ cell precursors. To determine what proteins the GLHs bind, C. elegans cDNA libraries were screened by the yeast two-hybrid method, using GLHs as bait. Three interacting proteins, CSN-5, KGB-1, and ZYX-1, were identified and further characterized. GST pull-down assays independently established that these proteins bind GLHs. CSN-5 is closely related to the subunit 5 protein of COP9 signalosomes, conserved multiprotein complexes of plants and animals. RNA interference (RNAi) with csn-5 results in sterile worms with small gonads and no oocytes, a defect essentially identical to that produced by RNAi with a combination of glh-1 and glh-4. KGB-1 is a putative JNK MAP kinase that GLHs bind. A kgb-1 deletion strain has a temperature-sensitive, sterile phenotype characterized by the absence of mature oocytes and the presence of trapped, immature oocytes that have undergone endoreplication. ZYX-1 is a LIM domain protein most like vertebrate Zyxin, a cytoskeletal adaptor protein. In C. elegans, while zyx-1 appears to be a single copy gene, neither RNAi depletion nor a zyx-1 deletion strain results in an obvious phenotype. These three conserved proteins are the first members in each of their families reported to associate with germline helicases. Similar to the loss of GLH-1 and GLH-4, loss of either CSN-5 or KGB-1 causes oogenesis to cease, but does not affect the initial assembly of P granules.

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“…One case has been reported in which Jab1 interacted with the GAL4 BD domain during YTH screening (Nordgard et al 2001). Thus one step can be added in which the prey library is screened using the empty bait vector (with no cloned genes); this may help to find the false positives that interact with the GAL4-BD only.…”

Section: Discussionmentioning

confidence: 99%

Development of a high-throughput yeast two-hybrid screening system to study protein-protein interactions in plants

Fang

Xue

et al. 2002

Mol Gen Genomics

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We have developed a high-throughput yeast two-hybrid screening system (HTP-YTH) that incorporates yeast gap-repair cloning, multiple positive ( ADE2, HIS3, lacZ) and negative ( URA3-based) selection schemes to reduce the incidence of negative and false positive clones, and automation of laboratory procedures to increase throughput. This HTP-YTH system has been applied to the study of protein-protein interactions that are involved in rice defense signal transduction pathways. More than 100 genes involved in plant defense responses were selected from DuPont's rice expressed sequence tag (EST) databases as baits for HTP-YTH screening. Results from YTH screening of eight of these rice genes are presented in this paper. Not only have we identified known protein-protein interactions, but we have also discovered novel interactions, which may ultimately reveal the regulatory network of host defense signal transduction pathways. We have demonstrated that our HTP-YTH method can be used to map protein-protein interaction networks and signal transduction pathways in any system. In combination with other approaches, such efficient YTH screens can help us systemically to study the functions of known and unknown genes in the genomics era.

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JAB1/CSN5 interacts with the GAL4 DNA binding domain: A note of caution about two-hybrid interactions

Cited by 29 publications

References 28 publications

Geminiviruses Subvert Ubiquitination by Altering CSN-Mediated Derubylation of SCF E3 Ligase Complexes and Inhibit Jasmonate Signaling inArabidopsis thaliana

Geminiviruses Subvert Ubiquitination by Altering CSN-Mediated Derubylation of SCF E3 Ligase Complexes and Inhibit Jasmonate Signaling inArabidopsis thaliana

The GLH Proteins, Caenorhabditis elegans P Granule Components, Associate with CSN-5 and KGB-1, Proteins Necessary for Fertility, and with ZYX-1, a Predicted Cytoskeletal Protein

Development of a high-throughput yeast two-hybrid screening system to study protein-protein interactions in plants

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