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PubMedDate not supplied

Myeloid differentiation (MyD) primary response genes in hematopoiesis.

Myeloid differentiation (MyD) primary response and growth arrest DNA-damage (Gadd) genes comprise a set of overlapping genes, including known (IRF-1, EGR-1, Jun) and novel (MyD88, Gadd45a MyD118/Gadd45b, GADD45g, MyD116/Gadd34) genes, that have been cloned by virtue of being coordinately induced upon the onset of terminal myeloid differentiation. This review delineates the role MyD genes were found to play in blood cell development, where they function as positive regulators of terminal differentiation, lineage specific blood cell development, and control of blood cell homeostasis, including growth inhibition and apoptosis.

Animals↗
PubMed2002

Myeloid differentiation (MyD)/growth arrest DNA damage (GADD) genes in tumor suppression, immunity and inflammation.

Myeloid differentiation (MyD) primary response and growth arrest DNA damage (Gadd) genes comprise a set of overlapping genes, including known (IRF-1, EGR-1, Jun) and novel (MyD88, Gadd45alpha, MyD118/Gadd45beta, GADD45gamma, MyD116/ Gadd34) genes, that have been cloned by virtue of being co-ordinately induced upon the onset of terminal myeloid differentiation and following exposure of cells to stress stimuli. In recent years it has become evident that MyD/Gadd play a role in blood cell development, where they function as positive regulators of terminal differentiation, lineage-specific blood cell development and control of blood cell homeostasis, including growth inhibition and apoptosis. MyD/Gadd are also involved in inflammatory responses to invading micro-organisms, and response to environmental stress and physiological stress, such as hypoxia, which results in ischemic tissue damage. An intricate network of interactions among MyD/GADD genes and gene products appears to control their diverse functions. Deregulated growth, increased cell survival, compromised differentiation and deficiencies in DNA repair are hallmarks of malignancy and its progression. Thus, the role MyD/Gadd play in negative growth control, including cell cycle arrest and apoptosis, and in DNA repair, make them attractive molecular targets for tumor suppression. The role MyD/Gadd play in innate immunity and host response to hypoxia also make these genes and gene products attractive molecular targets to treat immunity and inflammation disorders, such as septic shock and ischemic tissue damage.

Animals↗
PubMed2002

Myeloid differentiation (MyD) primary response genes in hematopoiesis.

Myeloid Differentiation (MyD) primary response and Growth Arrest DNA-Damage (Gadd) genes comprise a set of overlapping genes, including known (IRF-1, EGR-1, Jun) and novel (MyD88, Gadd45alpha MyD118/Gadd45beta, GADD45gamma, MyD116/Gadd34) genes, that have been cloned by virtue of there being co-ordinately induced upon the onset of terminal myeloid differentiation. This review delineates the role MyD genes play in blood cell development, where they function as positive regulators of terminal differentiation, lineage specific blood cell development and control of blood cell homeostasis, including growth inhibition and apoptosis.

Animals↗
PubMed1988

Nucleotide sequence of chicken myb proto-oncogene promoter region: detection of an evolutionarily conserved element.

The nucleotide sequence of the chicken myb proto-oncogene putative promoter region was determined and compared with the corresponding sequence of the mouse c-myb gene (1). 118 bp upstream from the initiation codon suggested by Gerondakis and Bishop (2) for the chicken c-myb protein, a 124-bp-long conserved element was found (92% identity in chicken and mouse sequences). Sequences homologous to this element were detected on Southern blots of restricted genomic DNAs from mouse, man, lizard, frog, and carp. No hybridization was observed with Drosophila, yeast, or Escherichia coli DNA. In human DNA, sequences homologous to this element were located at the 5' end of the c-myb gene, i.e. in the same position as in the chicken and mouse genes. Several lines of evidence suggest that the element is not a coding exon of a gene overlapping the c-myb gene. It may be of importance that one of the DNase I-sensitive sites and several c-myb mRNA cap sites localized recently in the mouse c-myb gene (3,4) lie within this region. It is suggested that this evolutionarily conserved element is involved in the regulation of myb proto-oncogene expression in vertebrates.

Animals↗
PubMed2001

Cell wall core galactofuran synthesis is essential for growth of mycobacteria.

The mycobacterial cell wall core consists of an outer lipid (mycolic acid) layer attached to peptidoglycan via a galactofuranosyl-containing polysaccharide, arabinogalactan. This structural arrangement strongly suggests that galactofuranosyl residues are essential for the growth and viability of mycobacteria. Galactofuranosyl residues are formed in nature by a ring contraction of UDP-galactopyranose to UDP-galactofuranose catalyzed by the enzyme UDP-galactopyranose mutase (Glf). In Mycobacterium tuberculosis the glf gene overlaps, by 1 nucleotide, a gene, Rv3808c, that has been shown to encode a galactofuranosyl transferase. We demonstrate here that glf can be knocked out in Mycobacterium smegmatis by allelic replacement only in the presence of two rescue plasmids carrying functional copies of glf and Rv3808c. The glf rescue plasmid was designed with a temperature-sensitive origin of replication and the M. smegmatis glf knockout mutant is unable to grow at the higher temperature at which the glf-containing rescue plasmid is lost. In a separate experiment, the Rv3808c rescue plasmid was designed with a temperature-sensitive origin of replication and the glf-bearing plasmid was designed with a normal original of replication; this strain was also unable to grow at the nonpermissive temperature. Thus, both glf and Rv3808c are essential for growth. These findings and the fact that galactofuranosyl residues are not found in humans supports the development of UDP-galactopyranose mutase and galactofuranosyl transferase as important targets for the development of new antituberculosis drugs.

Bacterial Proteins↗
PubMed1993

The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks.

All mammalian hepadnaviruses possess a gene, termed X, that encodes a protein capable of transactivating virus gene expression. The X gene overlaps the polymerase and precore genes as well as two newly identified open reading frames (ORFs) termed ORF5 and ORF6. In this investigation, we examined whether ORF5, ORF6, and the X gene were important for the replication of woodchuck hepatitis virus (WHV) in susceptible woodchucks. First, we investigated whether proteins were produced from ORF5 and ORF6 by in vitro translation of appropriate viral transcripts, searched for antibodies against the putative proteins in the sera of animals infected with wild-type virus, and looked for an antisense WHV transcript, necessary for expression of a protein from ORF6, in the livers of acutely or chronically infected woodchucks. All such experiments yielded negative results. Next, we used oligonucleotide-directed mutagenesis to introduce termination codons into ORF5 and ORF6 at two locations within each ORF. Adult woodchucks in groups of three were transfected with one of the four mutant genomes. All of these woodchucks developed WHV infections that were indistinguishable from those of animals transfected with the wild-type WHV recombinant. Polymerase chain reaction amplification and direct DNA sequencing confirmed that reversion of the mutants to a wild-type genotype did not occur. Taken together, these data indicate that ORF5 and ORF6 are not essential for virus replication and are unlikely to represent authentic genes. Finally, we generated five WHV X-gene mutants that either removed the initiation codon for protein synthesis or truncated the carboxyl terminus of the protein by 3, 16, 31, or 52 amino acids. Groups of three adult woodchucks were transfected with one of the five X-gene mutants. Only the mutant that possessed an X gene lacking 3 amino acids from the carboxyl terminus was capable of replication within the 6-month time frame of the experiment. In contrast, all seven woodchucks transfected with wild-type WHV DNA developed markers consistent with viral infection. Thus, it is likely (P < 0.01) that the WHV X gene is important for virus replication in the natural host.

Amino Acid Sequence↗
PubMed1978

The peptide chain elongation factor genes tufA and fus of Escherichia coli are intimately related physically.

Recent work from several laboratories has established the following points about the synthesis of the polypeptide chain elongation factors Tu and G in Escherichia coli. (i) Elongation factor Tu is the product of duplicate, highly conserved genes, tufA and tufB, which are widely separate parts of the chromosome. (ii) The molar concentration of this factor is considerably higher than that of elongation factor G which is encoded by the fus gene. (iii) Although the tufA and fus genes are close together and can be co-transcribed in the direction from fus to tufA, the tufA gene product is synthesized at several times the rate of the fus gene product. In an attempt to understand what mechanism(s) could account for the differential expression of the tufA and fus genes, we sought to obtain more precise information on the physical relationship of these genes. By examining heteroduplexes between restriction endonuclease-generated fragments of DNA containing the tufA, fus, and tufB genes, we have demonstrated that the fus and tufA genes are intimately related physically in one of two possible arrangements. Either the NH2-terminal region of the tufA gene is contiguous with the COOH-terminal region of the fus gene or the beginning of the tufA gene overlaps part of the fus gene. These results mean that if the tufA gene is always co-transcribed with the fus gene, then some mechanism must allow the tufA portion of the transcript to be translated more often than the fus gene portion of the transcript.

DNA, Bacterial↗
PubMed2001

Evidence for a new hepatitis C virus antigen encoded in an overlapping reading frame.

Many viruses have overlapping genes and/or regions in which a nucleic acid signal is embedded in a coding sequence. To search for dual-use regions in the hepatitis C virus (HCV), we developed a facile computer-based sequence analysis method to map dual-use regions in coding sequences. Eight diverse full-length HCV RNA and polyprotein sequences were aligned and analyzed. A cluster of unusually conserved synonymous codons was found in the core-encoding region, indicating a potential overlapping open reading frame (ORF). Four peptides (A1, A2, A3, and A4) representing this alternate reading frame protein (ARFP), two others from the HCV core protein, and one from bovine serum albumin (BSA) were conjugated to BSA and used in western blots to test sera for specific antibodies from 100 chronic HCV patients, 44 healthy controls, and 60 patients with non-HCV liver disease. At a 1:20,000 dilution, specific IgGs to three of the four ARFP peptides were detected in chronic HCV sera. Reactivity to either the A1 or A3 peptides (both ARFP derived) was significantly associated with chronic HCV infection, when compared to non-HCV liver disease serum samples (10/100 versus 1/60; p < 0.025). Antibodies to A4 were not detected in any serum sample. Our western blot assays confirmed the presence of specific antibodies to a new HCV antigen encoded, at least in part, in an alternate reading frame (ARF) overlapping the core-encoding region. Because this novel HCV protein stimulates specific immune responses, it has potential value in diagnostic tests and as a component of vaccines. This protein is predicted to be highly basic and may play a role in HCV replication, pathogenesis, and carcinogenesis.

Alternative Splicing↗
PubMed1992

Translational frameshifting mediated by a viral sequence in plant cells.

It has been proposed that the polymerase gene of barley yellow dwarf virus and related viruses is expressed by a ribosomal frameshift event during translation. The 5' end of this gene overlaps with the 3' end of an upstream gene that is in a different reading frame. The region of overlap is similar to sequences in retro- and coronaviruses that are known to express their polymerase genes by frameshifting. This overlap region includes a "shifty" heptanucleotide, followed by a highly structured region that may contain a pseudoknot. Sequences of 115 or 144 base pairs that span this region from barley yellow dwarf virus (PAV serotype) genomic RNA were introduced into a plasmid, so that a reporter gene could be expressed in plant cells only if a minus one (-1) frameshift event occurred. Frameshifting was detected at a rate of approximately 1%. This frameshifting was abolished when the stop codon at the 3' end of the upstream open reading frame was deleted. A sequence expected to form a strong stem-loop immediately upstream of the frameshift site was unnecessary for frameshifting, and initiation at AUG codons within the stem-loop appeared to be inhibited. Like viruses that infect hosts in other kingdoms, plant viruses also can induce frameshifting in translation of their genes.

Amino Acid Sequence↗
PubMed1991

Tryptophan gene cluster of Methanobacterium thermoautotrophicum Marburg: molecular cloning and nucleotide sequence of a putative trpEGCFBAD operon.

A recombinant cosmid carrying the Methanobacterium thermoautotrophicum Marburg trp genes was selected by complementation of Escherichia coli trp mutations. A 7.3-kb fragment of the cloned archaeal DNA was sequenced. It contained the seven trp genes, arranged adjacent to each other in the order trpEGCFBAD. No gene fusions were observed. The trp genes were organized in an operonlike structure, with four short (5- to 56-bp) intergenic regions and two overlapping genes. There was no indication for an open reading frame encoding a leader peptide in the upstream region of trpE. The gene order observed in the M. thermoautotrophicum trp operon was different from all known arrangements of the trp genes in archaea, bacteria, and eucarya. The encoded sequences of the Methanobacterium Trp proteins were similar in size to their bacterial and eucaryal counterparts, and all of them contained the segments of highly similar or invariant amino acid residues recognized in the Trp enzymes from bacteria and eucarya. The TrpE, TrpG, TrpC, TrpA, and TrpD proteins were 30 to 50% identical to those from representatives of other species. Significantly less sequence conservation (18 to 30%) was observed for TrpF, and TrpB exhibited a high degree of identity (50 to 62%) to the sequences of representatives of the three domains. With the exception of TrpB, the beta subunit of tryptophan synthase, tryptophan was absent from all Trp polypeptides.

Amino Acid Sequence↗
PubMed1991

Expression strategy of the potato virus X triple gene block.

The mode of expression of the overlapping genes of the triple block positioned internally in potato virus X (PVX) RNA was examined. The results of In vitro translation of synthetic RNA transcripts and natural PVX-specific methylmercuric hydroxide-denatured dsRNAs suggest that the 25K protein is expressed as a single translation product of the 2.1 kb subgenomic (sg) RNA and that both the 12K and 8K proteins are expressed from the same 1.4 kb sgRNA.

Base Sequence↗
PubMed2003

[Construction of a chimeric SEA-hPLAP-1 cDNA with gene splicing by overlap extension].

OBJECTIVE: To construct a chimeric SEA-hPLAP-1 cDNA with gene splicing by overlap extension. METHODS: The SEA gene and a DNA fragment encoding the signal for GPI-anchor attachment of hPLAP -1 were amplified by PCR. The two amplified gene sequence was annealed to form a chimeric GPI- anchored SEA molecule with gene splicing by overlap extension. The resulting chimera was cloned in pGEM-T vector and verified by sequencing analysis. RESULT: A chimeric SEA-hPLAP-1 cDNA was successfully constructed with gene splicing by overlap extension. CONCLUSION: Gene splicing by overlap extension is a successful specific PCR technique for gene recombination.

Alkaline Phosphatase↗
PubMed1984

Transformation mediated by the SV40 T antigens: separation of the overlapping SV40 early genes with a retroviral vector.

A murine retroviral vector has been used to separate physically the overlapping genes encoded by SV40. This minimal retroviral vector contains LTRs and other cis-acting signals required for infectious RNA virus propagation. We placed the SV40 early region within this DNA and after transfection of cells producing helper Moloney murine leukemia virus, SV40 retroviruses (MV40) could be rescued. Cytoplasmic spliced large T and small t transcripts, as well as unspliced transcripts, are packaged into virions with equal efficiency. Pure SV40 large T retroviruses can be cloned from these heterogeneous virus stocks by secondary transformation of rodent cells. The large T retrovirus stocks morphologically transform primary or established mouse and rat lines with high efficiency. There is little difference in transformation either by agar assay or focus formation between retroviruses carrying both SV40 genes or large T alone. We present quantitative data that demonstrate that abortive transformation of rodent cells by SV40, transient expression of the transformed phenotype after infection, is not manifested by MV40. Thus abortive transformation is not the result of a weakly dominant transforming gene, but rather of the normally inefficient mode of integration and early gene expression of SV40 upon infection of rodent cells.

Antigens, Viral, Tumor↗
PubMed1996

Genomic organization and sequence conservation in type I Trichomonas vaginalis viruses.

To reveal the genetic conservation of type I Trichomonas vaginalis viruses (TVV) we cloned and sequenced the 4.6-kb ds RNA of a TVV-T5 isolate for comparison with the cDNA sequence of a related TVV-T1 ds RNA. Analogous to TVV-T1, the TVV-T5 ds RNA also contains an upstream capsid protein gene overlapped with a downstream RNA-dependent RNA polymerase (RDRP) gene by a +1 reading frame shift. A conserved ribosomal slippage heptamer (C CUU UUU) was found within the consensus 14-nt overlap, and the context of the sequence surrounding the heptamer suggests a potential ribosomal frameshifting in the biosynthesis of RDRP from the initiation of capsid protein either through two consecutive -1 shifts or a +1 shift.

Animals↗
PubMed1991

Genetic and biochemical analysis of the aspartokinase from Corynebacterium glutamicum.

The lysC/asd gene cluster of Corynebacterium glutamicum ATCC 13032 was cloned and sequenced. The lysC locus coding for aspartokinase consists of two in-frame overlapping genes, lysC alpha encoding a protein of 421 amino acids (Mr 44,300) and lysC beta encoding a protein of 172 amino acids (Mr 18,600). The C. glutamicum aspartokinase was purified and found to contain two proteins of Mr 47,000 and Mr 18,000. A C. glutamicum mutant expressing a feedback-resistant aspartokinase was shown to be changed in a single base pair of the lysC beta gene, leading to an amino acid exchange in the beta-subunit of the aspartokinase. In addition, the identified mutation was found to be responsible for the enhanced expression of the asd gene located downstream of lysC.

Amino Acid Sequence↗
PubMed2006

Autism and environmental genomics.

Autism spectrum disorders (ASD) are defined by behavior and diagnosed by clinical history and observation but have no biomarkers and are presumably, etiologically and biologically heterogeneous. Given brain abnormalities and high monozygotic concordance, ASDs have been framed as neurobiologically based and highly genetic, which has shaped the research agenda and in particular criteria for choosing candidate ASD genes. Genetic studies to date have not uncovered genes of strong effect, but a move toward "genetic complexity" at the neurobiological level may not suffice, as evidence of systemic abnormalities (e.g. gastrointestinal and immune), increasing rates and less than 100% monozygotic concordance support a more inclusive reframing of autism as a multisystem disorder with genetic influence and environmental contributors. We review this evidence and also use a bioinformatic approach to explore the possibility that "environmentally responsive genes" not specifically associated with the nervous system, but potentially associated with systemic changes in autism, have not hitherto received sufficient attention in autism genetics investigations. We overlapped genes from NIEHS Environmental Genome Project, the Comparative Toxicogenomics Database, and the SeattleSNPs database of genes relevant to the human immune and inflammatory response with linkage regions identified in published autism genome scans. We identified 135 genes in overlap regions, of which 56 had never previously been studied in relation to autism and 47 had functional SNPs (in coding regions). Both our review and the bioinformatics exercise support the expansion of criteria for evaluating the relevance of genes to autism risk to include genes related to systemic impact and environmental responsiveness. This review also suggests the utility of environmental genomic resources in highlighting the potential relevance of particular genes within linkage regions. Environmental responsiveness and systems impacts consistent with system-wide findings in autism are thus supported as important considerations in identifying the numerous and complex modes of gene-environment interaction in autism.

Animals↗
PubMed2005

Transcription in the prpC-yloQ region in Bacillus subtilis.

The prpC and prkC genes encode two proteins with opposing activities, eukaryotic-like protein phosphatase and protein kinase involved in the sporulation and biofilm formation processes. The prpC gene overlaps 3 bp of the prkC gene; prkC is followed by the essential gene, yloQ. The organisation of the prpC, prkC and yloQ genes is conserved among several Gram-positive bacteria. In this work, we have found two promoters which function within the region of these genes and we also described conditions in which these promoters become activated. One promoter appears to be located upstream of prpC resulting in transcription of both prpC and prkC, as well as the yloQ gene. A second promoter is located 7 bp upstream of the start codon of yloQ. The yloQ promoter was activated during ethanol stress, but was sigmaB-independent. We also showed that prpC, prkC and yloQ genes form an operon.

Bacillus subtilis↗
PubMed1989

Dorsal expression of the Drosophila z600 gene during early embryogenesis.

The Drosophila z600 gene is a member of an overlapping gene cluster located in the 71CD interval of chromosome 3. We have used several approaches to study the spatial and temporal expression of z600 during embryogenesis. Northern analysis shows that z600 is zygotically expressed, with gene transcripts accumulating transiently during early embryo-genesis. The localization of z600 transcripts in embryo sections by in situ hybridization reveals a dynamic pattern of RNA distribution. z600 RNA is distributed throughout the embryo during the midsyncytial blastoderm stage, becomes concentrated dorsally and posteriorly during cellularization, and persists in the dorsal-most region of the embryo during gastrulation and germ band extension. z600 transcript accumulation is altered in dorsalized or ventralized mutant embryos, suggesting that z600 is directly or indirectly regulated by genes specifying the dorsal-ventral pattern in the embryo. An analysis of germ line transformants harboring a z600-lacZ gene fusion demonstrates that a 276-bp DNA region contains the sequences responsible for dorsal activation, but not ventral repression, of z600 gene expression during early embryogenesis.

Animals↗