New ligands of the Cellular Retinoic Acid-Binding Protein 2 (CRABP2) suggest a role for this protein in chromatin remodeling

Retinoic acid (RA) regulates the transcription of a series of genes involved in cell proliferation, differentiation and apoptosis by binding to the RA Receptor (RAR) and Retinoid X Receptor (RXR) heterodimers. The cellular retinoic acid-binding protein 2 (CRABP2) is involved in the transport of RA from the cytosol to specific RA receptors in the nucleus, acting as a coactivator of nuclear retinoid receptors. In order to have a better understanding of the role of CRABP2 in RA signaling, we used the yeast two-hybrid system as a tool for the identification of physical protein-protein interactions. Twenty-three putative CRABP2-interacting proteins were identified by screening in the presence of RA, five of which are related to transcription regulation or, more specifically, to the process of chromatin remodeling: t-complex 1 (TCP1); H3 histone, family 3A (H3F3A); H3 histone, family 3B (H3F3B); β-tubulin (TUBB) and SR-related CTD-associated factor 1 (SCAF1). These results suggest a more direct role for CRABP2 in chromatin remodeling and may be a starting point for the elucidation of the fine-tuning control of transcription by RA receptors...

Papel da proteína Mx1 na resposta a interferons e no processo neoplásico / Role of Mx1 protein in the response to interferons and neoplastic process

A proteína Mx1 é codificada por um gene induzido por interferon e compartilha a organização de seus domínios, a capacidade de homo-oligomerização e associação com membranas com as grandes dinaminas GTPases. A proteína Mx1 está envolvida na resposta contra um grande número de vírus de RNA, como aqueles pertencentes à família Buniavírus e o vírus influenza. Curiosamente, o gene MX1 foi encontrado como silenciado por metilação em diversos processos neoplásicos, incluindo carcinomas de cabeça e pescoço de células escamosas. Neste cenário, o silenciamento gênico de MX1 está associado à imortalização de uma série de linhagens celulares neoplásicas. Assim, Mx1 se destaca como uma das principais proteínas envolvidas nas respostas imunes induzidas por interferon e também desempenha um importante papel no controle do ciclo celular. Aqui discutimos os aspectos funcionais da proteína Mx1 abordando sua atividade antiviral, organização estrutural, envolvimento com neoplasias e, principalmente, os aspectos funcionais obtidos pela determinação de seus parceiros celulares.

The Mx1 protein is encoded by an interferon-induced gene and shares domain organization, homo-oligomerization capacity and membrane association with the large dynamin-like GTPases. The Mx1 protein is involved in the response to a large number of RNA viruses, such as the bunyavirus family and the influenza virus. Interestingly, it has also been found as a methylation-silenced gene in several types of neoplasm, including head and neck squamous cell carcinoma. In this scenario, MX1 gene silencing is associated with immortalization in several neoplastic cell lines. Thus, Mx1 stands out as one of the key proteins involved in interferon-induced immune response and also plays an important role in cell cycle control. Here we discuss some of the functions of the Mx1 protein, including its antiviral activity, protein folding and involvement in neoplasia, as well as those revealed by investigating its cellular partners.

Use of a synthetic lethal screen to identify genes related to TIF51A in Saccharomyces cerevisiae

The putative eukaryotic translation initiation factor 5A (eIF5A) is an essential protein for cell viability and the only cellular protein known to contain the unusual amino acid residue hypusine. eIF5A has been implicated in translation initiation, cell proliferation, nucleocytoplasmic transport, mRNA decay, and actin polarization, but the precise biological function of this protein is not clear. However, eIF5A was recently shown to be directly involved with the translational machinery. A screen for synthetic lethal mutations was carried out with one of the temperature-sensitive alleles of TIF51A (tif51A-3) to identify factors that functionally interact with eIF5A and revealed the essential gene YPT1. This gene encodes a small GTPase, a member of the rab family involved with secretion, acting in the vesicular trafficking between endoplasmatic reticulum and the Golgi. Thus, the synthetic lethality between TIF51A and YPT1 may reveal the connection between translation and the polarized distribution of membrane components, suggesting that these proteins work together in the cell to guarantee proper protein synthesis and secretion necessary for correct bud formation during G1/S transition. Future studies will investigate the functional interaction between eIF5A and Ypt1 in order to clarify this involvement of eIF5A with vesicular trafficking.

eIF5A: uma proteína essencial para a viabilidade celular cuja função permanece obscura / eIF5A: an essential protein for cell viability whose function remains obscure

O provável fator de início de tradução 5A (eIF5A) é uma proteína abundante e altamente conservada em todos os organismos eucarióticos observados e também está presente em arquebactérias. eIF5A é essencial para aviabilidade celular e esse fator é a única proteína descrita que contém o resíduo de aminoácido hipusina. Em Saccharomyces cerevisiae, eIF5A é expressa em condições aeróbicas pelo gene TIF51A. Apesar de eIF5A ser conhecida há quase 30 anos, a sua função biológica ainda é obscura. Este artigo revisa os estudos de caracterização funcional de eIF5A, evidenciando como esse fator foi envolvido com diferentes etapas do metabolismo de RNA mensageiro (mRNA), como o início de tradução, o transporte nucleocitoplasmático e o decaimento de RNA mensageiro. Ainda, estudos que evidenciaram o envolvimento de eIF5A com a proliferação celular e progressão no ciclo celular também foram abordados. Finalmente, esse artigo apresenta os resultados recentes dos experimentos que colocam eIF5A novamente no cenário da tradução. Novos experimentos serão necessários para definir o papel desempenhado por eIF5A na maquinaria de tradução.

Molecular genetic approach to cell proliferation control and neoplasia

A number of gene products involved in the control of cell proliferation fall into one of two classes: oncogenes and tumor suppressor genes. The same gene products have also been associated with malignant growth (tumors) caused by radiation, chemicals and tumor viruses. Here we describe our attempts to elucidate the molecular mechanisms underlying polyomavirus-induced cell transformation and the anti-tumor activity of glucocorticoid hormones. Wild type and mutant polyomavirus middle T (MT) overexpressing cell lines, generated with retroviral vector constructs, were used to investigate the role played by peptide growth factor primary response genes (fos, jun, myc, JE, KC) in viral transformation and to map the transduction pathway of the mitogenic signal of MT. Overexpression of MT leads to increased AP-1 (Fos/Jun) transcriptional complex activity. Transformation defective mutant analysis allowed the identification of sites in the MT molecule that are crucial for this activity. Two different approaches were used to investigate the molecular basis for glucocorticoids anti-tumor activity, namely: blind cloning of cDNAs and analysis of growth control genes in C6 glioma cell variants that are either hypersensitive (C6/ST1) or unresponsive to glucocorticoids (C6/P7). Four different glucocorticoid-regulated cDNA sequences were isolated using differential hybridization. A number of differentially expressed sequences were isolated from glucocorticoid-treated C6/ST1 cells by differential display (DDRT-PCR) and are currently being characterized. Expression of known growth control genes in C6/ST1 cells allowed the identification of important candidates for glucocorticoid hormone targets.