Topics

Characterization of a novel separase-interacting protein and candidate new securin, Eip1p, in the fungal pathogen .

08:00 EDT 14th August 2019 | BioPortfolio

Summary of "Characterization of a novel separase-interacting protein and candidate new securin, Eip1p, in the fungal pathogen ."

Proper chromosome segregation is crucial for maintaining genomic stability, and dependent on separase, a conserved and essential cohesin protease. Securins are key regulators of separases, but remain elusive in many organisms due to sequence divergence. Here, we demonstrate that the separase homologue Esp1p in the ascomycete , an important pathogen of humans, is essential for chromosome segregation. However, lacks a sequence homologue of securins found in model ascomycetes. We sought a functional homologue through identifying Esp1p interacting factors. Affinity purification of Esp1p and mass spectrometry revealed Eip1p/Orf19.955p, an uncharacterized protein specific to species. Functional analyses demonstrated that Eip1p is important for chromosome segregation but not essential, and modulated in an APC-dependent manner, similar to securins. Eip1p is strongly enriched in response to MMS or HU treatment, and its depletion partially suppresses an MMS or HU-induced metaphase block. Further, Eip1p depletion reduces Mcd1p/Scc1p, a cohesin subunit and separase target. Thus, Eip1p may function as a securin. However, other defects in Eip1p-depleted cells suggest additional roles. Overall, the results introduce a candidate new securin, provide an approach for identifying these divergent proteins, reveal a putative anti-fungal therapeutic target, and highlight variations in mitotic regulation in eukaryotes. Movie S1 Movie S1 Time-lapse imaging of Htb1p-GFP demonstrating normal chromosome segregation in cells expressing . Strain SS44() was incubated in inducing medium for 5 h, followed by mounting on an 8-well μSlide in inducing medium. Images were captured every 5 min for 180 min. Movie S2 Movie S2 Time-lapse imaging of Htb1p-GFP in an Eip1p-depleted cell demonstrating uneven segregation of chromosomes within the mother cell. Strain SS44 () wasincubated in repressing medium for 5 h, followed by mounting on an 8-well μSlide in repressing medium. Images were captured every 5 min for 180 min. Movie S3 Movie S3 Time-lapse imaging of Htb1p-GFP in Eip1p-depleted cells demonstrating abnormal segregation of chromosomes or abnormal nuclear translocation between the mother and daughter cells. Strain SS44 () wasincubated in repressing medium for 5 h, followed by mounting on an 8-well μSlide in repressing medium. Images were captured every 5 min for 180 min. Movie S4 Movie S4 Time-lapse imaging of Tub2p-GFP demonstrating normal spindle assembly and disassembly during mitosis in cells expressing . Strain SS38 () was incubated in inducing medium for 5 h, followed by mounting on an 8-well μSlide in inducing medium. Images were captured every 5 min for 180 min. Movie S5 Movie S5 Time-lapse imaging of Tub2p-GFP in an Eip1p-depleted cell demonstrating maintenance and oscillations of an elongated spindle. Strain SS38 () wasincubated in repressing medium for 5 h, followed by mounting on an 8-well μSlide in repressing medium. Images were captured every 5 min for 180 min. Movie S6 Movie S6 Time-lapse imaging of Tub2p-GFP in an Eip1p-depleted cell demonstrating oscillations of spindle pole bodies between the mother and daughter cells. Strain SS38 () wasincubated in repressing medium for 5 h, followed by mounting on an 8-well μSlide in repressing medium. Images were captured every 5 min for 180 min.

Affiliation

Journal Details

This article was published in the following journal.

Name: Molecular biology of the cell
ISSN: 1939-4586
Pages: mbcE18110696

Links

DeepDyve research library

PubMed Articles [14937 Associated PubMed Articles listed on BioPortfolio]

Glycan analysis for protein therapeutics.

Glycosylation can be a critical quality attribute for protein therapeutics due to its extensive impact on product safety and efficacy. Glycan characterization is important in the process of protein dr...

Decrypting protein surfaces by combining evolution, geometry and molecular docking.

The growing body of experimental and computational data describing how proteins interact with each other has emphasized the multiplicity of protein interactions and the complexity underlying protein s...

Multicomponent Yeast Two-Hybrid System: Applications to Study Protein-Protein Interactions in SMC Complexes.

Analysis of protein-protein interactions (PPI) is key for the understanding of most protein assemblies including structural maintenance of chromosomes (SMC) complexes. SMC complexes are composed of SM...

Genome-wide association study and protein network analysis for understanding candidate genes involved in root development at the rapeseed seedling stage.

Root system is essential for plants to absorb water and nutrients. The root related traits are complex quantitative traits and regulated by genetic control. Here, we used two association mapping panel...

Circulating aryl hydrocarbon receptor-interacting protein (AIP) is independent of GH secretion.

Aryl hydrocarbon receptor-interacting protein (AIP) is evolutionarily conserved and expressed widely throughout the organism. Loss-of-function AIP mutations predispose to young-onset pituitary adenoma...

Clinical Trials [3784 Associated Clinical Trials listed on BioPortfolio]

PRO-STATE:Search for a Protein Profile Corresponding to Fast-Developing Lesions and Characterization of Implicated Proteins in Prostate Carcinoma

The main objective of this study is to realise serum protein profiles for each patient undergoing a prostate biopsy and to identify relevant proteins.

Double-Blind Study of Safety and Immunogenicity of Two Candidate Malaria Vaccines in Gabonese Children

GSK Biologicals is developing a number of candidate malaria vaccines for the routine immunization of infants and children living in malaria-endemic areas. The candidate vaccines are design...

Positional Cloning of the Gene(s) Responsible for Alagille Syndrome

The goal of the project is to identify and clone the gene(s) responsible for the Alagille Syndrome (AGS) by a positional cloning approach. The first step towards this goal is to define th...

MP0250 DARPin® Protein Plus Osimertinib in Patients With EGFR-mutated NSCLC

The purpose of this study is to assess the anti-tumor efficacy, safety, tolerability, pharmacokinetics (PK), immunogenicity and biological activity of the MP0250 DARPin® drug candidate in...

Genetic Epidemiology of Lipoprotein-Lipid Levels

To determine the contribution of polymorphic variation in candidate genes involved in lipid metabolism in determining quantitative lipoprotein-lipid levels and cardiovascular risk factors ...

Medical and Biotech [MESH] Definitions

Securin is involved in the control of the metaphase-anaphase transition during MITOSIS. It promotes the onset of anaphase by blocking SEPARASE function and preventing proteolysis of cohesin and separation of sister CHROMATIDS. Overexpression of securin is associated with NEOPLASTIC CELL TRANSFORMATION and tumor formation.

Separase is a caspase-like cysteine protease, which plays a central role in triggering ANAPHASE by cleaving the SCC1/RAD21 subunit of the cohesin complex. Cohesin holds the sister CHROMATIDS together during METAPHASE and its cleavage results in chromosome segregation.

A family of serine-threonine kinases that plays a role in intracellular signal transduction by interacting with a variety of signaling adaptor proteins such as CRADD SIGNALING ADAPTOR PROTEIN; TNF RECEPTOR-ASSOCIATED FACTOR 2; and TNF RECEPTOR-ASSOCIATED DEATH DOMAIN PROTEIN. Although they were initially described as death domain-binding adaptor proteins, members of this family may contain other protein-binding domains such as those involving caspase activation and recruitment.

A member of the Bcl-2 protein family that reversibly binds MEMBRANES. It is a pro-apoptotic protein that is activated by caspase cleavage.

Techniques for measuring specific nucleic acid interaction with another nucleic acid or with a protein by digestion of the non-interacting nucleic acid by various nucleases. After all non-interacting regions are eliminated by nuclease digestion, the protected nucleic acid that remains is analyzed. DNA FOOTPRINTING utilizes this technique to analyze the DNA contact sites of DNA-BINDING PROTEINS.

Quick Search


DeepDyve research library

Relevant Topic

Bioinformatics
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...


Searches Linking to this Article