Prenylated proteins contain a covalently linked cholesterol intermediate near their carboxyl-termini. Maturation of most prenylated involves proteolytic removal the last three amino acids. Two genes in Saccharomyces cerevisiae , RCE1 and AFC1 were identified that appear to be responsible for this processing. The Afc1 protein is zinc protease participates processing yeast -factor mating pheromone. Rce1 contributes both Ras -factor. Deletion resulted loss proteins. Disruption led defects...

Candida albicans is a ubiquitous opportunistic pathogen that the most prevalent cause of hospital-acquired fungal infections. In mammalian hosts, C. engulfed by phagocytes attack with DNA-damaging reactive oxygen species (ROS). Acetylation histone H3 lysine 56 (H3K56) fungal-specific acetyltransferase Rtt109 important for yeast model organisms to survive DNA damage and maintain genome integrity. To assess importance pathogenicity, we deleted predicted homolog in clinical isolate, SC5314....

The B10.Q/J strain of mice was serendipitously discovered to be highly susceptible infection by the intracellular protozoan parasite, Toxoplasma gondii but markedly resistant induction autoimmune arthritis. We have previously shown that phenotype is controlled a single recessive locus and associated with lymphocyte hyporesponsiveness IL-12. Using genetic approaches, we now localized chromosome 9 established its identity as Tyk2 , Janus kinase essential for IL-12 IFN-α/β cytokine signaling....

Many proteins that contain a carboxyl-terminal CaaX sequence motif, including Ras and yeast a-factor, undergo series of sequential posttranslational processing steps. Following the initial prenylation cysteine, three C-terminal amino acids are proteolytically removed, newly formed prenylcysteine is carboxymethylated. The specific comprise influence whether protein can be prenylated proteolyzed. In this study, we evaluated a-factor variants with all possible single acid substitutions at...

ROR family of nuclear receptor transcription factors forms nodes connecting metabolic and inflammatory signaling pathways. The RORα members the have intrinsic transcriptional activity they are involved in both activation repression a wide range genes. role control inflammation has been extensively studied using animal models but its function human cells is not as well understood. To address this shortcoming, we analyzed how shaping state macrophages. Using CRISPR-Cas9 system, deleted RORA...

Characterization of essential genes across the genome is fundamental to understanding cellular functions at a molecular level. While significant progress has been made in characterizing human and mouse models, relatively little known about porcine genome. Pigs are an important production species now emerging as valuable models for studying diseases due their physiological similarities humans. To map genome, we have developed novel genome-wide CRISPR knockout screening library (pGeCKO)...

Intrahepatic and extrahepatic metastases are common findings in hepatocellular carcinoma (HCC). Insulin-like growth factor 2 (IGF2) expression is frequently induced HCC, serum IGF2 levels correlate with the presence of metastases. Yet, role IGF-induced signaling dissemination HCC remains unclear. We have previously observed elevated tumors metastatic potential an mouse model. Here, we demonstrate that inhibition IGF2, or its receptor IGF1R, impairs migration invasion activities murine cells....

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related death worldwide. HCC patients frequently present with disease that has metastasized to other regions the liver, portal vein, lymph nodes, or lungs, poor prognoses. Therefore, model systems allow exploration molecular mechanisms underlying metastasis in this are greatly needed. We describe here metastatic generated after somatic introduction mouse polyoma virus middle T antigen mice liver-specific deletion Trp53 tumor...

Haemophilus influenzae efficiently colonizes and persists at the human nasopharyngeal mucosa, causing disease when it spreads to other sites. Nitric oxide (NO) represents a major antimicrobial defense deployed by host cells in locations colonized H. during pathogenesis that are likely vary oxygen levels. Formate-dependent nitrite reductase regulator (FNR) is an oxygen-sensitive several bacterial pathogens. We report fnr of required for anaerobic against exposure NO donors resist NO-dependent...

Host cells use metabolic signaling through the LXRα nuclear receptor to defend against Listeria monocytogenes infection. 25-Hydroxycholesterol is a natural ligand of LXRs that produced by enzyme cholesterol 25-hydroxylase (CH25H). We found expression Ch25h upregulated following L. infection in beta interferon (IFN-β)-dependent fashion. Moreover, increased promotes survival monocytogenes-infected and increases sensitivity host determined Cd5l, prosurvival gene, controlled CH25H. In addition,...

Abstract The transcriptional repressor B lymphocyte–induced maturation protein 1 (BLIMP1) is a master regulator of and T cell differentiation. To examine the role BLIMP1 in innate immunity, we used conditional knockout (CKO) Blimp1 myeloid cells found that CKO mice were protected from lethal infection induced by Listeria monocytogenes. Transcriptome analysis macrophages identified murine chemokine (C-C motif) ligand 8, CCL8, as direct target Blimp1-mediated repression these cells....

Abstract In eukaryotes small secreted peptides are often proteolytically cleaved from larger precursors. Saccharomyces cerevisiae multiple proteolytic processing steps required for production of mature 12-amino-acid a-factor its 36-amino-acid precursor. This study provides additional genetic data supporting a direct role Afc1p in cleavage the carboxyl-terminal tripeptide CAAX motif prenylated addition, had second that was independent of, and addition to, vivo. Using ubiquitin-a-factor...

Abstract Spt3 of Saccharomyces cerevisiae is required for the normal transcription many genes in vivo. Past studies have shown that both mating and sporulation, two events initiate when cells are at G1/START. We now show needed other begin G1/START, diploid filamentous growth haploid invasive growth. In addition, expression FLO11, a gene growth, although this defect not sole cause spt3Δ/spt3Δ defect. To extend our Spt3's role to pathogenic yeast Candida albicans, we identified C. albicans...

Abstract Epidemiological, clinical, and experimental approaches have convincingly demonstrated that host resistance to infection with intracellular pathogens is significantly influenced by genetic polymorphisms. Using a mouse model of virulent Mycobacterium tuberculosis (MTB), we previously identified the sst1 locus as determinant tuberculosis. In this study demonstrate susceptibility another pathogen, Listeria monocytogenes, also locus. The contribution anti-listerial immunity much greater...

Hepatic gluconeogenesis is required for maintaining blood glucose homeostasis; yet, in diabetes mellitus, this process unrestrained and a major contributor to fasting hyperglycemia. To date, the impacts of chromatin modifying enzymes landscape on are poorly understood. Through catalyzing removal methyl groups from specific lysine residues histone tail, demethylases modulate structure and, hence, gene expression. Here we perform an RNA interference screen against known identify H3 36 (H3K36)...

Abstract We isolated a mutant defective in C‐terminal farnesyl cysteine:carboxyl methyltransferase activity from screen for mutations causing ‐specific sterility. A genomic fragment was cloned yeast multi‐copy library that restored mating. Both the gene and sterile mutation were allelic to STE14 gene. ste14 ‐complementing 2·17 kb Bam HI subclone sequenced found encode 239 amino acid protein with molecular weight of 27,887 Daltons. The hydrophobicity profile reveals presence at least five...

Genetic makeup of the host plays a significant role in course and outcome infection. Inbred strains mice display wide range sensitivities to Listeria monocytogenes infection thus serve as good model for analysis effect genetic polymorphism. The L. is influenced by ability this bacterium induce expression interferon beta mRNA, encoded mouse Ifnb1 (interferon 1, fibroblast) gene. Mouse that lack components IFN signaling pathway are substantially more resistant We found macrophages from ByJ...

MULIBREY nanism which results from autosomal recessive mutations in TRIM37 impacts skeletal development, leading to growth delay with complications multiple organs. In this study, we employed a combined proteomics and qPCR screening approach investigate the molecular alterations CHON-002 cell line by comparing wild-type (WT) cells knockdown (KD) cells. Our proteomic analysis demonstrated that depletion predominantly affects expression of extracellular matrix proteins (ECM). Specifically,...

Protein prenyltransferases catalyze the covalent attachment of isoprenoid lipids (farnesyl or geranylgeranyl) to a cysteine near C terminus their substrates. This study explored specificity determinants for interactions between farnesyltransferase Saccharomyces cerevisiae and its protein A series substitutions at amino acid 149 β-subunit were tested in combination with C-terminal CaaX substrates Ras2p a-factor. Efficient prenylation was observed when oppositely charged acids present yeast...

Host susceptibility to infection is controlled in large measure by the genetic makeup of host. Spirochetes genus Borrelia include nearly 40 species vector-borne spirochetes that are capable infecting a wide range mammalian hosts, causing Lyme disease and relapsing fever. Relapsing fever associated with high-level bacteremia, as well hematologic manifestations, such thrombocytopenia (i.e., low platelet numbers) anemia. To facilitate studies control hermsii infection, we performed systematic...