Úloha evolučně konzervovaných proteinů BIR-1/Survivin a SKP-1 v regulaci genové exprese

Souhrn SKIP a BIR/Survivin jsou evolučně zachovalé proteiny. SKIP je známý transkripční a sestřihový kofaktor a BIR-1/Survivin reguluje bunečné dělení, genovou expresi a vývoj. Inaktivace SKP-1 a BIR-1 indukuje podobné vývojové fenotypy. K odhalení možných interakcí SKP-1 a BIR-1 jsme použili kvasinkový dvojhybridní systém a knihovnu kompletní mRNA C. elegans. Tyto experimenty identifikovaly částečně se překrývající kategorie proteinů jako interaktory proteinů SKP-1 a BIR-1. Identifikované interagující proteiny zahrnovaly ribozomální proteiny, transkripční faktory, translační faktory, cytoskeletální a motorové proteiny. Tyto výsledky naznačují jejich možnou účast v mnohočetných proteinových komplexech. Pomocí krátkodobé nadměrné exprese BIR-1 jsme sledovali účinek BIR-1 na proteom C. elegans v larválním stádiu L1. To způsobilo dramatickou změnu v celém proteomu což naznačuje, že BIR-1 má schopnost změnit chromatografický profil mnohočetných cílových proteinů včetně těch, které jsme již dříve identifikovali jako interagující proteiny v experimentach s kvasinkovým dvouhybridním systému. Výsledky jsme následně potvrdili pro RPS-3, RPL-5, myosin (non-muscle myosin) a TAC-1 (transkripční kofaktor a protein asociovaný s centrosomy). Tyto výsledky naznačují, že SKP-1 a BIR-1 jsou multifunkční proteiny, které jsou...SKIP and BIR/Survivin are evolutionarily conserved proteins. SKIP is a known transcription and splicing cofactor while BIR-1/Survivin regulates cell division, gene expression and development. Loss of function of C. elegans SKIP (SKP-1) and BIR-1 induces overlapping developmental phenotypes. In order to uncover the possible interactions of SKP-1 and BIR-1 on the protein level, we screened the complete C. elegans mRNA library using the yeast two-hybrid system. These experiments identified partially overlapping categories of proteins as SKP-1 and BIR-1 interactors. The interacting proteins included ribosomal proteins, transcription factors, translation factors and cytoskeletal and motor proteins suggesting involvement of the two studied proteins in multiple protein complexes. To visualize the effect of BIR-1 on the proteome of C. elegans we induced a short time pulse BIR-1 overexpression in synchronized L1 larvae. This led to a dramatic alteration of the whole proteome pattern indicating that BIR-1 alone has the capacity to alter the chromatographic profile of many target proteins including proteins found to be interactors in yeast two hybrid screens. The results were validated for ribosomal proteins RPS-3, RPL-5, non-muscle myosin and TAC-1, a transcription cofactor and a centrosome associated...Ústav buněčné biologie a patologie 1. LF UKInstitute of Cell Biology and Pathology First Faculty of Medicine Charles UniversityFirst Faculty of Medicine1. lékařská fakult

Quality of life in patients after radical orchiectomy

Testikulární nádory představují sice jen 1% všech maligních onemocnění, které postihují mužskou populaci, ale ve věkové skupině 20-44 let dosahuje jejich podíl 23%. Představují nejčastěji se vyskytující nádor bílé populace v této věkové skupině. Orchiektomie má zásadní význam jak v diagnostice tak v léčbě testikulárních nádorů. Je téměř nemožné zkoumat pacienty z pouhého hlediska důsledků radikální orchiektomie, jelikož jen malá skupina pacientů prodělá pouze tento operační zákrok. Téměř nedílnou součástí následné terapie je adjuvantní radioterapie a chemoterapie (které značně ovlivňují pacienta). Dále samotné nádorové onemocnění působí negativně na organizmus. Z těchto důvodů nelze na kvalitu života pacientů nahlížet pouze z hlediska radikální orchiektomie, ale musíme se na kvalitu života pacientů po orchiektomii dívat z komplexního pohledu testikulárních nádorů. Musíme zvážit důsledky i ostatních terapeutických modalit. Skupina pacientů, kteří přežili testikulární nádorové onemocnění (v anglickém jazyce-testicular cancer survivors-TCS) představují zajímavou skupinu ke sledovaní kvality života z několika důvodů. Léčba testikulárních nádorů je v 90% úspěšná a následně mají TCS stejnou predikovanou dobu dožití jako stejně staří muži ze zdravé populace. TCS mají také určitá specifika v otázkách fertility a...Although testicular tumors represent only 1% of all malignancies that affect the male population , but in the age group of 20-44 years reaching their share of 23%. They represent the most frequent tumor of the white population in that age group. Orchiectomy is critical both in diagnosis and in treatment testicular tumors . It is almost impossible to examine the patient merely from the point of view consequences of radical orchiectomy , since only a small group of patients will experience only this surgery . Almost an integral part of the subsequent therapy is adjuvant radiotherapy and chemotherapy ( which significantly affect patient). Furthermore itself cancer has a negative effect on the organism. For these reasons, the quality of life of patients seen only in terms of radical orchiectomy , but We have the quality of life of patients after orchiectomy look of a comprehensive Looking testicular tumors . We must consider the consequences of other therapeutic modalities. The group of patients who survived testicular cancer ( in English - testicular - cancer survivors TCS) are an interesting group to monitor the quality of life for several reasons. Treatment of testicular tumors in 90 % successful and consequently have the same TCS predicted life expectancy as well old men from the healthy population. TCS...Urologická klinikaDepartment of UrologyThird Faculty of Medicine3. lékařská fakult

The role of evolutionarily conserved proteins BIR-1/Survivin and SKP-1 in the regulation of gene expression

SKIP and BIR/Survivin are evolutionarily conserved proteins. SKIP is a known transcription and splicing cofactor while BIR-1/Survivin regulates cell division, gene expression and development. Loss of function of C. elegans SKIP (SKP-1) and BIR-1 induces overlapping developmental phenotypes. In order to uncover the possible interactions of SKP-1 and BIR-1 on the protein level, we screened the complete C. elegans mRNA library using the yeast two-hybrid system. These experiments identified partially overlapping categories of proteins as SKP-1 and BIR-1 interactors. The interacting proteins included ribosomal proteins, transcription factors, translation factors and cytoskeletal and motor proteins suggesting involvement of the two studied proteins in multiple protein complexes. To visualize the effect of BIR-1 on the proteome of C. elegans we induced a short time pulse BIR-1 overexpression in synchronized L1 larvae. This led to a dramatic alteration of the whole proteome pattern indicating that BIR-1 alone has the capacity to alter the chromatographic profile of many target proteins including proteins found to be interactors in yeast two hybrid screens. The results were validated for ribosomal proteins RPS-3, RPL-5, non-muscle myosin and TAC-1, a transcription cofactor and a centrosome associated..

Quality of life in patients after radical orchiectomy

Although testicular tumors represent only 1% of all malignancies that affect the male population , but in the age group of 20-44 years reaching their share of 23%. They represent the most frequent tumor of the white population in that age group. Orchiectomy is critical both in diagnosis and in treatment testicular tumors . It is almost impossible to examine the patient merely from the point of view consequences of radical orchiectomy , since only a small group of patients will experience only this surgery . Almost an integral part of the subsequent therapy is adjuvant radiotherapy and chemotherapy ( which significantly affect patient). Furthermore itself cancer has a negative effect on the organism. For these reasons, the quality of life of patients seen only in terms of radical orchiectomy , but We have the quality of life of patients after orchiectomy look of a comprehensive Looking testicular tumors . We must consider the consequences of other therapeutic modalities. The group of patients who survived testicular cancer ( in English - testicular - cancer survivors TCS) are an interesting group to monitor the quality of life for several reasons. Treatment of testicular tumors in 90 % successful and consequently have the same TCS predicted life expectancy as well old men from the healthy population. TCS..

Trichoplax adhaerens reveals a network of nuclear receptors sensitive to 9-cis-retinoic acid at the base of metazoan evolution

Trichoplax adhaerens, the only known species of Placozoa is likely to be closely related to an early metazoan that preceded branching of Cnidaria and Bilateria. This animal species is surprisingly well adapted to free life in the World Ocean inhabiting tidal costal zones of oceans and seas with warm to moderate temperatures and shallow waters. The genome of T. adhaerens (sp. Grell) includes four nuclear receptors, namely orthologue of RXR (NR2B), HNF4 (NR2A), COUP-TF (NR2F) and ERR (NR3B) that show a high degree of similarity with human orthologues. In the case of RXR, the sequence identity to human RXR alpha reaches 81% in the DNA binding domain and 70% in the ligand binding domain. We show that T. adhaerens RXR (TaRXR) binds 9-cis retinoic acid (9-cis-RA) with high affinity, as well as high specificity and that exposure of T. adhaerens to 9-cis-RA regulates the expression of the putative T. adhaerens orthologue of vertebrate L-malate-NADP+ oxidoreductase (EC 1.1.1.40) which in vertebrates is regulated by a heterodimer of RXR and thyroid hormone receptor. Treatment by 9-cis-RA alters the relative expression profile of T. adhaerens nuclear receptors, suggesting the existence of natural ligands. Keeping with this, algal food composition has a profound effect on T. adhaerens growth and appearance. We show that nanomolar concentrations of 9-cis-RA interfere with T. adhaerens growth response to specific algal food and causes growth arrest. Our results uncover an endocrine-like network of nuclear receptors sensitive to 9-cis-RA in T. adhaerens and support the existence of a ligand-sensitive network of nuclear receptors at the base of metazoan evolution

Molecular cloning and sequence of retinoid X receptor in the green crab Carcinus maenas: a possible role in female reproduction

Retinoid X receptor (RXR) belongs to an ancient superfamily of nuclear hormone receptors, and plays an important role in reproduction of vertebrates. However, the reproductive role of RXR has not been clarified in crustaceans. In this investigation, we first report the cloning of two alternative splice variants of RXR cDNA from green crab ovarian RNA. RXR mRNA levels were quantified in different vitellogenic stages of the crab hepatopancreas (HP) and ovary. The expression of RXR mRNA relative to the arginine kinase mRNA was significantly increased in the HP of vitellogenic crabs in a stage-dependent manner. The relative levels of RXR mRNA in the ovary were significantly lower in vitellogenic stage III crabs than in crabs in the other three stages. These data indicate that the HP and ovary of the crab are capable of expressing RXR, which may regulate, in part, vitellogenesis in the crab. We also examined the effects of methyl farnesoate (MF) and RXR-dsRNA treatments on vitellogenin and RXR gene expression. Vitellogenin and RXR mRNA levels in HP and ovarian fragments incubated in MF were significantly (

A two-step process in the emergence of neurogenesis

Cnidarians belong to the first phylum differentiating a nervous system, thus providing suitable model systems to trace the origins of neurogenesis. Indeed corals, sea anemones, jellyfish and hydra contract, swim and catch their food thanks to sophisticated nervous systems that share with bilaterians common neurophysiological mechanisms. However, cnidarian neuroanatomies are quite diverse, and reconstructing the urcnidarian nervous system is ambiguous. At least a series of characters recognized in all classes appear plesiomorphic: (1) the three cell types that build cnidarian nervous systems (sensory-motor cells, ganglionic neurons and mechanosensory cells called nematocytes or cnidocytes); (2) an organization of nerve nets and nerve rings [those working as annular central nervous system (CNS)]; (3) a neuronal conduction via neurotransmitters; (4) a larval anterior sensory organ required for metamorphosis; (5) a persisting neurogenesis in adulthood. By contrast, the origin of the larval and adult neural stem cells differs between hydrozoans and other cnidarians; the sensory organs (ocelli, lens-eyes, statocysts) are present in medusae but absent in anthozoans; the electrical neuroid conduction is restricted to hydrozoans. Evo-devo approaches might help reconstruct the neurogenic status of the last common cnidarian ancestor. In fact, recent genomic analyses show that if most components of the postsynaptic density predate metazoan origin, the bilaterian neurogenic gene families originated later, in basal metazoans or as eumetazoan novelties. Striking examples are the ParaHox Gsx, Pax, Six, COUP-TF and Twist-type regulators, which seemingly exert neurogenic functions in cnidarians, including eye differentiation, and support the view of a two-step process in the emergence of neurogenesis