scholarly journals The mod-A Suppressor of Nonallelic Heterokaryon Incompatibility in Podospora anserina Encodes a Proline-Rich Polypeptide Involved in Female Organ Formation

Genetics ◽  
1998 ◽  
Vol 149 (2) ◽  
pp. 915-926 ◽  
Author(s):  
Christian Barreau ◽  
Maya Iskandar ◽  
Gabriel Loubradou ◽  
Véronique Levallois ◽  
Joël Bégueret
Keyword(s):  
Protein Interactions ◽  
Cellular Differentiation ◽  
Growth Arrest ◽  
Podospora Anserina ◽  
The Self ◽  
Protein Protein Interactions ◽  
Vegetative Incompatibility ◽  
Binding Motifs ◽  
Reading Frame ◽  
Incompatibility Reaction

Abstract Vegetative incompatibility in fungi results from the control of heterokaryon formation by the genes present at het loci. Coexpression of antagonistic het genes in the same hyphae leads to a lethal process. In Podospora anserina, self-incompatible strains containing nonallelic incompatible genes in the same nucleus are inviable as the result of a growth arrest and a lytic process. Mutations in suppressor genes (mod genes) can restore the viability. These mod mutations also interfere with developmental processes, which suggests common steps between the incompatibility reaction and cellular differentiation. The mod-A locus, responsible for growth arrest in the self-incompatible strains, is also involved in the control of the development of female organs. The mod-A gene was isolated. An open reading frame 687 amino acids long was identified. The MOD-A-encoded polypeptide is rich in proline residues, which are clustered in a domain containing a motif that displays similarity to SH3-binding motifs, which are known to be involved in protein-protein interactions. Construction of a strain deleted for mod-A confirmed that the product of this gene involved in differentiation is a key regulator of growth arrest associated with vegetative incompatibility.

A guide-tag system controlling client enrichment into Y15 peptide-based granules for an in-cell protein recruitment assay

2021 ◽  
Author(s):  
Takayuki Miki ◽  
Masahiro Hashimoto ◽  
Taichi Nakai ◽  
Hisakazu Mihara
Keyword(s):  
Protein Interactions ◽  
Living Cells ◽  
The Self ◽  
Cell Protein ◽  
Protein Protein Interactions ◽  
Self Assembling ◽  
Client Proteins ◽  
Protein Recruitment ◽  
Peptide Tag

A series of guide-tags that can control the enrichment of client proteins into artificial scaffolds constituted by the self-assembling Y15 peptide tag facilitates the analysis of protein–protein interactions in living cells.

scholarly journals ArabidopsisImmunophilin-like TWD1 Functionally Interacts with Vacuolar ABC Transporters

2004 ◽  
Vol 15 (7) ◽  
pp. 3393-3405 ◽  
Author(s):  
Markus Geisler ◽  
Marjolaine Girin ◽  
Sabine Brandt ◽  
Vincent Vincenzetti ◽  
Sonia Plaza ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Abc Transporters ◽  
Loss Of Function ◽  
Protein Protein Interactions ◽  
Binding Motifs ◽  
Calmodulin Binding ◽  
C Terminus ◽  
Domain Mapping ◽  
Tetratricopeptide Repeat Domain

Previously, the immunophilin-like protein TWD1 from Arabidopsis has been demonstrated to interact with the ABC transporters AtPGP1 and its closest homologue, AtPGP19. Physiological and biochemical investigation of pgp1/pgp19 and of twd1 plants suggested a regulatory role of TWD1 on AtPGP1/AtPGP19 transport activities. To further understand the dramatic pleiotropic phenotype that is caused by loss-of-function mutation of the TWD1 gene, we were interested in other TWD1 interacting proteins. AtMRP1, a multidrug resistance-associated (MRP/ABCC)-like ABC transporter, has been isolated in a yeast two-hybrid screen. We demonstrate molecular interaction between TWD1 and ABC transporters AtMRP1 and its closest homologue, AtMRP2. Unlike AtPGP1, AtMRP1 binds to the C-terminal tetratricopeptide repeat domain of TWD1, which is well known to mediate protein-protein interactions. Domain mapping proved that TWD1 binds to a motif of AtMRP1 that resembles calmodulin-binding motifs; and calmodulin binding to the C-terminus of MRP1 was verified. By membrane fractionation and GFP-tagging, we localized AtMRP1 to the central vacuolar membrane and the TWD1-AtMRP1 complex was verified in vivo by coimmunoprecipitation. We were able to demonstrate that TWD1 binds to isolated vacuoles and has a significant impact on the uptake of metolachlor-GS and estradiol-β-glucuronide, well-known substrates of vacuolar transporters AtMRP1 and AtMRP2.

scholarly journals Evidence for Multimerization of Neu Proteins Involved in Polysialic Acid Synthesis in Escherichia coliK1 Using Improved LexA-Based Vectors

2000 ◽  
Vol 182 (18) ◽  
pp. 5267-5270 ◽  
Author(s):  
Dayle A. Daines ◽  
Richard P. Silver
Keyword(s):  
Protein Interactions ◽  
Polysialic Acid ◽  
Genetic System ◽  
Acid Synthesis ◽  
Full Length ◽  
Protein Protein Interactions ◽  
Protein Fragment ◽  
Reading Frame ◽  
E Coli

ABSTRACT Recently, M. Dmitrova et al. (Mol. Gen. Genet. 257:205–212, 1998) described a LexA-based genetic system to monitor protein-protein interactions in an Escherichia coli background. However, the plasmids used in this system, pMS604 and pDP804, were not readily amenable for general use. In this report, we describe modifications of both plasmids that allow fragments of DNA to be fused to either vector in any reading frame. Homodimerization and heterodimerization of full-length proteins involved in polysialic acid synthesis in E. coli K1, as well as heterodimerization between a full-length protein and a protein fragment, demonstrate the usefulness of the modified plasmids for investigating bacterial protein-protein interactions in vivo.

scholarly journals Regulation of Gene Expression During the Vegetative Incompatibility Reaction in Podospora anserina: Characterization of Three Induced Genes

Genetics ◽  
1998 ◽  
Vol 150 (2) ◽  
pp. 633-641
Author(s):  
Nathalie Bourges ◽  
Alexis Groppi ◽  
Christian Barreau ◽  
Corinne Clavé ◽  
Joël Bégueret
Keyword(s):  
Gene Expression ◽  
Expression Profiles ◽  
Regulation Of Gene Expression ◽  
Podospora Anserina ◽  
In Vitro Translation ◽  
Vegetative Incompatibility ◽  
Two Dimensional Gel Electrophoresis ◽  
Small Proteins ◽  
Mrna Content ◽  
Incompatibility Reaction

Abstract Vegetative incompatibility in fungi limits the formation of viable heterokaryons. It results from the coexpression of incompatible genes in the heterokaryotic cells and leads to a cell death reaction. In Podospora anserina, a modification of gene expression takes place during this reaction, including a strong decrease of total RNA synthesis and the appearance of a new set of proteins. Using in vitro translation of mRNA and separation of protein products by two-dimensional gel electrophoresis, we have shown that the mRNA content of cells is qualitatively modified during the progress of the incompatibility reaction. Thus, gene expression during vegetative incompatibility is regulated, at least in part, by variation of the mRNA content of specific genes. A subtractive cDNA library enriched in sequences preferentially expressed during incompatibility was constructed. This library was used to identify genomic loci corresponding to genes whose mRNA is induced during incompatibility. Three such genes were characterized and named idi genes for genes induced during incompatibility. Their expression profiles suggest that they may be involved in different steps of the incompatibility reaction. The putative IDI proteins encoded by these genes are small proteins with signal peptides. IDI-2 protein is a cysteine-rich protein. IDI-2 and IDI-3 proteins display some similarity in a tryptophan-rich region.

scholarly journals De novocoiled-coil peptides as scaffolds for disrupting protein–protein interactions

2018 ◽  
Vol 9 (39) ◽  
pp. 7656-7665 ◽  
Author(s):  
Jordan M. Fletcher ◽  
Katherine A. Horner ◽  
Gail J. Bartlett ◽  
Guto G. Rhys ◽  
Andrew J. Wilson ◽  
...  
Keyword(s):  
Protein Binding ◽  
Protein Interactions ◽  
Coiled Coils ◽  
Protein Protein Interactions ◽  
Binding Motifs ◽  
Helical Protein

Homo- and hetero-dimeric coiled coils as scaffolds for the presentation of α-helical protein-binding motifs.

A mutation outside the two zinc fingers of ADR1 can suppress defects in either finger

1992 ◽  
Vol 12 (12) ◽  
pp. 5758-5767
Author(s):  
S Camier ◽  
N Kacherovsky ◽  
E T Young
Keyword(s):  
Dna Binding ◽  
Protein Interactions ◽  
Zinc Fingers ◽  
Wild Type ◽  
Protein Protein Interactions ◽  
Site Mutation ◽  
Binding Motifs ◽  
Amino Terminal ◽  
Sequence Homologies ◽  
In The Wild

A second-site mutation that restored DNA binding to ADR1 mutants altered at different positions in the two zinc fingers was identified. This mutation (called IS1) was a conservative change of arginine 91 to lysine in a region amino terminal to the two zinc fingers and known from previous experiments to be necessary for DNA binding. IS1 increased binding to the UAS1 sequence two- to sevenfold for various ADR1 mutants and twofold for wild-type ADR1. The change of arginine 91 to glycine decreased binding twofold, suggesting that this arginine is involved in DNA binding in the wild-type protein. The increase in binding by IS1 did not involve protein-protein interactions between the two ADR1 monomers, nor did it require the presence of the sequences flanking UAS1. However, the effect of IS1 was influenced by the sequence of the first finger, suggesting that interactions between the region amino terminal to the fingers and the fingers themselves could exist. A model for the role of the amino-terminal region based on these results and sequence homologies with other DNA-binding motifs is proposed.

Purification, characterization, and cDNA cloning of an AU-rich element RNA-binding protein, AUF1

1993 ◽  
Vol 13 (12) ◽  
pp. 7652-7665 ◽  
Author(s):  
W Zhang ◽  
B J Wagner ◽  
K Ehrenman ◽  
A W Schaefer ◽  
C T DeMaria ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Rna Binding ◽  
K562 Cells ◽  
Mrna Degradation ◽  
Protein Protein Interactions ◽  
Rna Recognition Motifs ◽  
Reading Frame ◽  
A Cell ◽  
Recognition Motifs ◽  
Macrophage Colony

The degradation of some proto-oncogene and lymphokine mRNAs is controlled in part by an AU-rich element (ARE) in the 3' untranslated region. It was shown previously (G. Brewer, Mol. Cell. Biol. 11:2460-2466, 1991) that two polypeptides (37 and 40 kDa) copurified with fractions of a 130,000 x g postribosomal supernatant (S130) from K562 cells that selectively accelerated degradation of c-myc mRNA in a cell-free decay system. These polypeptides bound specifically to the c-myc and granulocyte-macrophage colony-stimulating factor 3' UTRs, suggesting they are in part responsible for selective mRNA degradation. In the present work, we have purified the RNA-binding component of this mRNA degradation activity, which we refer to as AUF1. Using antisera specific for these polypeptides, we demonstrate that the 37- and 40-kDa polypeptides are immunologically cross-reactive and that both polypeptides are phosphorylated and can be found in a complex(s) with other polypeptides. Immunologically related polypeptides are found in both the nucleus and the cytoplasm. The antibodies were also used to clone a cDNA for the 37-kDa polypeptide. This cDNA contains an open reading frame predicted to produce a protein with several features, including two RNA recognition motifs and domains that potentially mediate protein-protein interactions. These results provide further support for a role of this protein in mediating ARE-directed mRNA degradation.

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