Teneurins in development and disease

The teneurins are a novel type II transmembrane protein family originally discovered in Drosophila and highly conserved between invertebrates and vertebrates. Studies in invertebrates suggest important functions for the teneurins in many processes during development. However, still very little is known about the biological function and mechanism of action of the vertebrate teneurin family, which consists of four paralogs called teneurin-1 to -4. In the first part of my thesis, I analyzed the expression pattern and signaling mechanism of teneurin-1 during chick development. Teneurin-1 was prominently expressed in specific regions of the brain, and often complementary to teneurin-2. The presence of teneurin-1 and -2 in interconnected regions of the brain implies a role for teneurins in the establishment of appropriate neuronal connectivity. Using a novel antibody recognizing the teneurin-1 intracellular domain (ICD), N-terminal processing products were detected and nuclear staining was observed in specific neurons and tissues. This provides evidence for our working hypothesis, according to which teneurins can be processed by a mechanism called regulated intramembrane proteolysis, resulting in the release and nuclear translocation of the ICD. Similar results were obtained for teneurin-4 during chick development. In the second part of my thesis, I investigated the implication of teneurins in two human diseases. X-linked mental retardation: Teneurin-1 was analyzed as an X-linked mental retardation (XLMR) candidate gene in 23 XLMR patients. No mutation that is likely to cause the disease was found the coding region or splice sites of the teneurin-1 gene in these patients. Brain tumors: Teneurin-4 was found to be upregulated in a microarray analysis of human brain tumors including astrocytomas, oligodendrogliomas and glioblastomas (GBMs). The overexpression of teneurin-4 was confirmed on protein level in brain tumor lysates. Immunohistochemistry (IHC) revealed strong staining around tumors cells in some brain tumors whereas in others teneurin-4 restricted to blood vessels

Integrative genomic analysis reveals widespread enhancer regulation by p53 in response to DNA damage

The tumor suppressor p53 has been studied extensively as a direct transcriptional activator of protein-coding genes. Recent studies, however, have shed light on novel regulatory functions of p53 within noncoding regions of the genome. Here, we use a systematic approach that integrates transcriptome-wide expression analysis, genome-wide p53 binding profiles and chromatin state maps to characterize the global regulatory roles of p53 in response to DNA damage. Notably, our approach identified conserved features of the p53 network in both human and mouse primary fibroblast models. In addition to known p53 targets, we identify many previously unappreciated mRNAs and long noncoding RNAs that are regulated by p53. Moreover, we find that p53 binding occurs predominantly within enhancers in both human and mouse model systems. The ability to modulate enhancer activity offers an additional layer of complexity to the p53 network and greatly expands the diversity of genomic elements directly regulated by p53

A Large Intergenic Noncoding RNA Induced by p53 Mediates Global Gene Repression in the p53 Response

Recently, more than 1000 large intergenic noncoding RNAs (lincRNAs) have been reported. These RNAs are evolutionarily conserved in mammalian genomes and thus presumably function in diverse biological processes. Here, we report the identification of lincRNAs that are regulated by p53. One of these lincRNAs (lincRNA-p21) serves as a repressor in p53-dependent transcriptional responses. Inhibition of lincRNA-p21 affects the expression of hundreds of gene targets enriched for genes normally repressed by p53. The observed transcriptional repression by lincRNA-p21 is mediated through the physical association with hnRNP-K. This interaction is required for proper genomic localization of hnRNP-K at repressed genes and regulation of p53 mediates apoptosis. We propose a model whereby transcription factors activate lincRNAs that serve as key repressors by physically associating with repressive complexes and modulate their localization to sets of previously active genes.National Institutes of Health (U.S.) (New Innovator Award)Smith Family FoundationDamon Runyon Cancer Research FoundationSearle Scholars ProgramNational Institutes of Health (U.S.) (1R01CA119176-01

Missense mutations in TENM4, a regulator of axon guidance and central myelination, cause essential tremor

Essential tremor (ET) is a common movement disorder with an estimated prevalence of 5% of the population aged over 65 years. In spite of intensive efforts, the genetic architecture of ET remains unknown. We used a combination of whole-exome sequencing and targeted resequencing in three ET families. In vitro and in vivo experiments in oligodendrocyte precursor cells and zebrafish were performed to test our findings. Whole-exome sequencing revealed a missense mutation in TENM4 segregating in an autosomal-dominant fashion in an ET family. Subsequent targeted resequencing of TENM4 led to the discovery of two novel missense mutations. Not only did these two mutations segregate with ET in two additional families, but we also observed significant over transmission of pathogenic TENM4 alleles across the three families. Consistent with a dominant mode of inheritance, in vitro analysis in oligodendrocyte precursor cells showed that mutant proteins mislocalize. Finally, expression of human mRNA harboring any of three patient mutations in zebrafish embryos induced defects in axon guidance, confirming a dominant-negative mode of action for these mutations. Our genetic and functional data, which is corroborated by the existence of a Tenm4 knockout mouse displaying an ET phenotype, implicates TENM4 in ET. Together with previous studies of TENM4 in model organisms, our studies intimate that processes regulating myelination in the central nervous system and axon guidance might be significant contributors to the genetic burden of this disorde

Caspase-11 activation requires lysis of pathogen-containing vacuoles by IFN-induced GTPases

Lipopolysaccharide from Gram-negative bacteria is sensed in the host cell cytoplasm by a non-canonical inflammasome pathway that ultimately results in caspase-11 activation and cell death. In mouse macrophages, activation of this pathway requires the production of type-I interferons, indicating that interferon-induced genes have a critical role in initiating this pathway. Here we report that a cluster of small interferon-inducible GTPases, the so-called guanylate-binding proteins, is required for the full activity of the non-canonical caspase-11 inflammasome during infections with vacuolar Gram-negative bacteria. We show that guanylate-binding proteins are recruited to intracellular bacterial pathogens and are necessary to induce the lysis of the pathogen-containing vacuole. Lysis of the vacuole releases bacteria into the cytosol, thus allowing the detection of their lipopolysaccharide by a yet unknown lipopolysaccharide sensor. Moreover, recognition of the lysed vacuole by the danger sensor galectin-8 initiates the uptake of bacteria into autophagosomes, which results in a reduction of caspase-11 activation. These results indicate that host-mediated lysis of pathogen-containing vacuoles is an essential immune function and is necessary for efficient recognition of pathogens by inflammasome complexes in the cytosol