scholarly journals Multiple interactions are involved in a highly specific association of the Mod(mdg4)-67.2 isoform with the Su(Hw) sites in Drosophila

Open Biology ◽  
2017 ◽  
Vol 7 (10) ◽  
pp. 170150 ◽  
Author(s):  
Larisa Melnikova ◽  
Margarita Kostyuchenko ◽  
Varvara Molodina ◽  
Alexander Parshikov ◽  
Pavel Georgiev ◽  
...  
Keyword(s):  
Protein Complexes ◽  
Specific Interaction ◽  
Btb Domain ◽  
Rich Domain ◽  
Blocking Activity ◽  
Transgenic Lines ◽  
Enhancer Blocking ◽  
Specific Association ◽  
Protein Variants ◽  
Multiple Interactions

The best-studied Drosophila insulator complex consists of two BTB-containing proteins, the Mod(mdg4)-67.2 isoform and CP190, which are recruited to the chromatin through interactions with the DNA-binding Su(Hw) protein. It was shown previously that Mod(mdg4)-67.2 is critical for the enhancer-blocking activity of the Su(Hw) insulators and it differs from more than 30 other Mod(mdg4) isoforms by the C-terminal domain required for a specific interaction with Su(Hw) only. The mechanism of the highly specific association between Mod(mdg4)-67.2 and Su(Hw) is not well understood. Therefore, we have performed a detailed analysis of domains involved in the interaction of Mod(mdg4)-67.2 with Su(Hw) and CP190. We found that the N-terminal region of Su(Hw) interacts with the glutamine-rich domain common to all the Mod(mdg4) isoforms. The unique C-terminal part of Mod(mdg4)-67.2 contains the Su(Hw)-interacting domain and the FLYWCH domain that facilitates a specific association between Mod(mdg4)-67.2 and the CP190/Su(Hw) complex. Finally, interaction between the BTB domain of Mod(mdg4)-67.2 and the M domain of CP190 has been demonstrated. By using transgenic lines expressing different protein variants, we have shown that all the newly identified interactions are to a greater or lesser extent redundant, which increases the reliability in the formation of the protein complexes.

Stable and specific association between the yeast recombination and DNA repair proteins RAD1 and RAD10 in vitro

1992 ◽  
Vol 12 (7) ◽  
pp. 3041-3049
Author(s):  
L Bardwell ◽  
A J Cooper ◽  
E C Friedberg
Keyword(s):  
Dna Repair ◽  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Specific Interaction ◽  
Mitotic Recombination ◽  
Nucleotide Excision ◽  
Specific Association ◽  
Recombination Pathway ◽  
Cloned Gene

The RAD1 and RAD10 genes of Saccharomyces cerevisiae are two of at least seven genes which are known to be required for damage-specific recognition and/or damage-specific incision of DNA during nucleotide excision repair. RAD1 and RAD10 are also involved in a specialized mitotic recombination pathway. We have previously reported the purification of the RAD10 protein to homogeneity (L. Bardwell, H. Burtscher, W. A. Weiss, C. M. Nicolet, and E. C. Friedberg, Biochemistry 29:3119-3126, 1990). In the present studies we show that the RAD1 protein, produced by in vitro transcription and translation of the cloned gene, specifically coimmunoprecipitates with the RAD10 protein translated in vitro or purified from yeast. Conversely, in vitro-translated RAD10 protein specifically coimmunoprecipitates with the RAD1 protein. The sites of this stable and specific interaction have been mapped to the C-terminal regions of both polypeptides. This portion of RAD10 protein is evolutionarily conserved. These results are the first biochemical evidence of a specific association between any eukaryotic proteins genetically identified as belonging to a recombination or DNA repair pathway and suggest that the RAD1 and RAD10 proteins act at the same or consecutive biochemical steps in both nucleotide excision repair and mitotic recombination.

scholarly journals Chicken Y-box proteins chk-YB-1b and chk-YB-2 repress translation by sequence-specific interaction with single-stranded RNA

2000 ◽  
Vol 348 (2) ◽  
pp. 297-305 ◽  
Author(s):  
Shivalingappa K. SWAMYNATHAN ◽  
Ashok NAMBIAR ◽  
Ramareddy V. GUNTAKA
Keyword(s):  
Transcriptional Regulation ◽  
Translational Regulation ◽  
Rna Binding ◽  
Wide Spectrum ◽  
Specific Interaction ◽  
Regulation Of Gene Expression ◽  
Large Family ◽  
Long Terminal Repeats ◽  
Rich Domain

Y-Box proteins comprise a large family of multifunctional proteins with a wide spectrum of activities in both transcription and translational regulation of gene expression. Earlier, we have reported on the involvement of chk-YB-2 in transcriptional regulation of Rous sarcoma virus long terminal repeats and the involvement of chk-YB-1b in transcriptional regulation of alpha1(I) collagen genes. Here, we have investigated the potential role of chk-YB-2 and chk-YB-1b in RNA metabolism. We report that chk-YB-2 and chk-YB-1b are localized predominantly in the cytoplasm and that they both can bind single-stranded RNA in a sequence-specific and reversible manner. Well-conserved cold-shock domain, N-terminal proline-rich domain and the alternating clusters of acidic and basic amino acids located in the C-terminal ends of these two proteins were all found to be necessary for their RNA-binding ability. Further, we demonstrate that these two proteins inhibit translation in vitro and that binding to RNA is required for this inhibition. The significance of these results is discussed.

scholarly journals 3. Safety Assessment of SIN LVs Harboring Chromatin Insulators in the Sensitive Cdkn2a-/- In Vivo Genotoxicity Assay Show Enhancer-Blocking Activity of Specific Insulator Sequences

2015 ◽  
Vol 23 ◽  
pp. S2
Author(s):  
Monica Volpin ◽  
Andrea Calabria ◽  
Daniela Cesana ◽  
Erika Tenderini ◽  
Fabrizio Benedicenti ◽  
...  
Keyword(s):  
Safety Assessment ◽  
Blocking Activity ◽  
Enhancer Blocking

scholarly journals Native Tandem and Ion Mobility Mass Spectrometry Highlight Structural and Modular Similarities in Clustered-Regularly-Interspaced Shot-Palindromic-Repeats (CRISPR)-associated Protein Complexes From Escherichia coli and Pseudomonas aeruginosa

2012 ◽  
Vol 11 (11) ◽  
pp. 1430-1441 ◽  
Author(s):  
Esther van Duijn ◽  
Ioana M. Barbu ◽  
Arjan Barendregt ◽  
Matthijs M. Jore ◽  
Blake Wiedenheft ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mass Spectrometry ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Acquired Resistance ◽  
Native Mass Spectrometry ◽  
Ion Mobility Mass Spectrometry ◽  
E Coli ◽  
Ribonucleoprotein Complexes ◽  
Specific Association

The CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) immune system of bacteria and archaea provides acquired resistance against viruses and plasmids, by a strategy analogous to RNA-interference. Key components of the defense system are ribonucleoprotein complexes, the composition of which appears highly variable in different CRISPR/Cas subtypes. Previous studies combined mass spectrometry, electron microscopy, and small angle x-ray scattering to demonstrate that the E. coli Cascade complex (405 kDa) and the P. aeruginosa Csy-complex (350 kDa) are similar in that they share a central spiral-shaped hexameric structure, flanked by associating proteins and one CRISPR RNA. Recently, a cryo-electron microscopy structure of Cascade revealed that the CRISPR RNA molecule resides in a groove of the hexameric backbone. For both complexes we here describe the use of native mass spectrometry in combination with ion mobility mass spectrometry to assign a stable core surrounded by more loosely associated modules. Via computational modeling subcomplex structures were proposed that relate to the experimental IMMS data. Despite the absence of obvious sequence homology between several subunits, detailed analysis of sub-complexes strongly suggests analogy between subunits of the two complexes. Probing the specific association of E. coli Cascade/crRNA to its complementary DNA target reveals a conformational change. All together these findings provide relevant new information about the potential assembly process of the two CRISPR-associated complexes.

scholarly journals Insights on small-molecules inhibitors and protein-protein interactions of the T4SS

2014 ◽  
Vol 70 (a1) ◽  
pp. C582-C582
Author(s):  
Ingrid Um Nlend ◽  
Christian Baron
Keyword(s):  
Small Molecules ◽  
Virulence Factor ◽  
Protein Interactions ◽  
Protein Complexes ◽  
Cell Envelope ◽  
Gram Negative Bacteria ◽  
Protein Protein Interactions ◽  
Excellent Model ◽  
Assembly Factor ◽  
Multiple Interactions

ABSTRACT: Bacterial T4SS are complexes, constituted of 8 to 12 conserved proteins, used by many gram-negative bacteria for the translocation of proteins and DNA-protein complexes as well as for the transportation of DNA-protein complexes across their cell envelope. T4SS are excellent model targets for the development of antivirulence drugs as it is an essential virulence factor for many bacterial pathogens, such as Brucella. Antivirulence drugs that deprive the pathogen of its essential virulence factor, the T4SS, would constitute alternatives to or enhancements of current antibiotic treatment. VirB8, a conserved assembly factor in T4SS forms dimers that are very important for T4SS function in these pathogens. Due to its multiple interactions, VirB8 is an excellent model for the analysis of assembly factors but also a possible target for drugs that could target its protein–protein interactions, which would disarm bacteria by depriving them of their essential virulence functions.

scholarly journals Inferring interaction partners from protein sequences

2016 ◽  
Author(s):  
Anne-Florence Bitbol ◽  
Robert S. Dwyer ◽  
Lucy J. Colwell ◽  
Ned S. Wingreen
Keyword(s):  
Protein Interactions ◽  
Sequence Data ◽  
Protein Complexes ◽  
A Priori ◽  
Specific Interaction ◽  
Three Dimensional ◽  
Specific Protein ◽  
Protein Protein Interactions ◽  
Entropy Model ◽  
Interaction Partners

Specific protein-protein interactions are crucial in the cell, both to ensure the formation and stability of multi-protein complexes, and to enable signal transduction in various pathways. Functional interactions between proteins result in coevolution between the interaction partners. Hence, the sequences of interacting partners are correlated. Here we exploit these correlations to accurately identify which proteins are specific interaction partners from sequence data alone. Our general approach, which employs a pairwise maximum entropy model to infer direct couplings between residues, has been successfully used to predict the three-dimensional structures of proteins from sequences. Building on this approach, we introduce an iterative algorithm to predict specific interaction partners from among the members of two protein families. We assess the algorithm's performance on histidine kinases and response regulators from bacterial two-component signaling systems. The algorithm proves successful without any a priori knowledge of interaction partners, yielding a striking 0.93 true positive fraction on our complete dataset, and we uncover the origin of this surprising success. Finally, we discuss how our method could be used to predict novel protein-protein interactions.

scholarly journals A NYN domain protein directly interacts with DCP1 and is required for phyllotactic pattern in Arabidopsis

2021 ◽  
Author(s):  
Marlene Schiaffini ◽  
Clara Chicois ◽  
Aude Pouclet ◽  
Tiphaine Chartier ◽  
Elodie Ubrig ◽  
...  
Keyword(s):  
Protein Complexes ◽  
In Planta ◽  
Main Partner ◽  
Growth Defects ◽  
Phyllotactic Pattern ◽  
Protein Partner ◽  
Transgenic Lines ◽  
Domain Protein ◽  
The One ◽  
Transcriptomic Changes

ABSTRACTIn eukaryotes, general mRNA decay requires the decapping complex. The activity of this complex depends on its catalytic subunit, DCP2 and its interaction with decapping enhancers, including its main partner DCP1. Here, we report that in Arabidopsis, DCP1 also interacts with a NYN domain endoribonuclease, hence named DCP1-ASSOCIATED NYN ENDORIBONUCLEASE 1 (DNE1). Interestingly, we find DNE1 predominantly associated with DCP1 but not with DCP2 and reciprocally, suggesting the existence of two distinct protein complexes. We also show that the catalytic residues of DNE1 are required to repress the expression of mRNAs in planta upon transient expression. The overexpression of DNE1 in transgenic lines leads to growth defects and transcriptomic changes related to the one observed upon inactivation of the decapping complex. Finally, the combination of dne1 and dcp2 mutations, revealed a functional redundancy between DNE1 and DCP2 in controlling phyllotactic pattern formation in Arabidopsis. Our work identifies DNE1, a hitherto unknown DCP1 protein partner highly conserved in the plant kingdom and identifies its importance for developmental robustness.One-sentence summaryDNE1, a NYN domain protein interacts with the decapping activator DCP1 and, together with DCP2, specify phyllotactic patterns in Arabidopsis.

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