Failure to detect "cap" structures in mitochondrial DNA-coded poly(A)-containing RNA from HeLa cells

The structure of the 5'-termini has been investigated in mitochondrial DNA- coded poly(A)-containing RNA from HeLa cells. For this purpose, mitochondrial RNA isolated from cells labeled for 3 hours with [32P]orthophosphate in the presence of 20 µg/ml camptothecin, and selected for poly(A) content by two passages through oligo(dT)-cellulose, was digested either with the nuclease P1 or with a mixture of RNases: the digestion products were then fractionated by two-dimensional electrophoresis. No "cap" structures were detected under conditions where the presence of such structures in one out of five to ten RNA molecules would have been recognized. It is, therefore, likely that "cap" structures are completely absent in HeLa cell mitochondrial poly(A)-containing RNA

Violence is as American as Cherry Pie: Mass Incarceration and Juvenile Violence

The purpose of this chapter is to offer “food for thought” regarding an under researched area of juvenile violence causation: the possible connection between steadily increasing incarceration rates and steadily increasing incidents of school violence. Unfortunately, the negative, and sometimes violent, traits individuals develop while incarcerated are often brought out into their lives in society and personal lives, which often involve the raising of children. Research has documented the impacts that being incarcerated can have upon an individual. There is growing research supporting that these newly developed traits and behaviors can easily be imbedded in the children in which they have contact with upon release. The authors argue that we should not be surprised about the increases in juvenile violence given the constant flow of individuals in and out of American prisons. This is not to say that everyone who has served time will follow this path, but this is one area where actions and patterns of behavior which have been developed in one social environment can saturate another

The Impact of Mass Incarceration and Social Issues of Institutionalization on School Violence in Youths

In this chapter the authors examine the impact that mass incarceration has had upon children in the United States over the last several decades. Inherent in this examination is to discuss the impact of institutionalization on the propensity of committing violent acts by children. The authors also discuss the impact of these American phenomena on many aspects of juvenile delinquency and violence

Demographics of Quail Hunters in Oklahoma

We collected information from Oklahoma hunters during a telephone survey conducted in February 1997, to compare demographics of scaled quail (Callipepla squamata) and northern bobwhite quail (Colinus virginianus) hunters to other hunters. Eight hundred ninetytwo annual, 709 lifetime, and 376 senior citizen hunting license holders, stratified by county of residence, were interviewed. Respondents who hunted quail differed from other hunters by age group, age of first hunting experience, the proportion of their life (\u3e=16 years of age) they have owned an Oklahoma hunting license, education level, annual household income, access to a computer at work, access to a computer at home, and access to the Internet at work (P \u3c= 0.030). No difference (P \u3e= 0.219) was found between hunter types by residential location, hunting license type, proportion of life residing in Oklahoma, ethnic origin, and access to the Internet at home. This information can help wildlife managers better understand their quail hunting constituents and tailor agency programs to fit their needs

Life on Death Row

The purpose of this chapter is to examine the general conditions for those living on death row in the United States. This examination involves the various restrictions of freedom outside of cell, size and conditions in cells, allowances on personal property, and limits on food and recreation. Special attention is given to discussing the unique relationships that often develop between correctional officers and death row inmates. These relationships are explored through the phenomena of “Hustling” (i.e., any actions taken by an inmate to generate revenue, power, or control over their environment). This type of behavior often is used as a coping mechanism for death row inmates as they deal with their living environment and potential fate. An interesting aspect of this behavior is that it can also serve as a coping mechanism for correctional officers working through the unique demands placed on those working with condemned inmates

Drosophila Single-minded Represses Gene Transcription by Activating the Expression of Repressive Factors

AbstractThe Drosophila single-minded gene controls CNS midline cell development by both activating midline gene expression and repressing lateral CNS gene expression in the midline cells. The mechanism by which Single-minded represses transcription was examined using the ventral nervous system defective gene as a target gene. Transgenic-lacZ analysis of constructs containing fragments of the ventral nervous system defective regulatory region identified sequences required for lateral CNS transcription and midline repression. Elimination of Single-minded:Tango binding sites within the ventral nervous system defective gene did not affect midline repression. Mutants of Single-minded that removed the DNA binding and transcriptional activation regions abolished ventral nervous system defective repression, as well as transcriptional activation of other genes. The replacement of the Single-minded transcriptional activation region with a heterologous VP16 transcriptional activation region restored the ability of Single-minded to both activate and repress transcription. These results indicate that Single-minded indirectly represses transcription by activating the expression of repressive factors. Single-minded provides a model system for how regulatory proteins that act only as transcriptional activators can control lineage-specific transcription in both positive and negative modes

The Structure of Mammalian Genes: (1) Antibody Heavy Chain Variable Region Genes: Organization, Diversity, and Somatic Mutation. (2) Structure and Transcription of the DNA Encompassing the Origin of Replication of Human Mitochondrial DNA

This thesis describes two experimental systems utilized to study mammalian gene structure and expression: (1) antibody heavy chain variable region genes and (2) mitochondrial DNA. In order to study the organization and structure of antibody genes and the relative germline and somatic contributions towards antibody diversity, we have analyzed the germline genes encoding the murine immune response to phosphorylcholine. Molecular cloning studies were undertaken and conclusively show that there is only one germline VH gene segment encoding the immune response to phosphorylcholine. Protein sequencing work on monoclonal antibodies that bind phosphorylcholine reveals many different protein sequences related to one predominant sequence. We are able to conclude that these variant sequences are the result of somatic diversification operating on one germline gene segment. We are further able to show that this diversification is mutational and not recombinational. Finally, somatic mutation is correlated with the class of the antibody; IgG and IgA antibodies undergo somatic mutation, IgM antibodies do not. We have isolated and sequenced a family of four closely related VH gene segments designated V1, V3, V11 and V13. Their function varies: V1 encodes the immune response to phosphorylcholine, V3 is a pseudogene, V11 encodes the immune response to influenza hemagglutinin, and V13 has an unknown function but is not obviously a pseudogene. Structural analysis of recombinant clones containing this family of related VH gene segments and other VH gene segments reveals several important points about the organization of VH gene segments. First, closely related VH gene segments can be clustered together within the VH gene locus. Second, the spacing distance between adjacent VH gene segments is variable; it may be as short as 5 kb and greater than 30 kb. Finally, the average spacing distance between VH gene segments is large, at least 23 kb. Assuming a minimum of 200 germline VH gene segments, the size of the VH gene locus may be greater than 5 million base pairs. The human mitochondrial genome is the second system that has been chosen to study gene structure and expression, and to accomplish this, we have applied both DNA and RNA sequencing technologies. We sequenced the DNA encompassing the origin of DNA replication and then localized the origin at the nucleotide level. The human mitochondrial origin of DNA replication shares structural characteristics with other known origins of DNA replication; in particular, the presence of extensive secondary structure in the form of a stem-loop structure. In order to precisely localize mitochondrial transcripts to the DNA, we developed techniques that allowed the isolation and sequencing of the 5'-ends of mitochondrial transcripts. This technology was utilized to precisely localize the 5'-end of the mitocohndrial 12S rRNA species 457 nucleotide pairs 5'-to the origin of DNA replication. Analysis of the DNA sequence in this region revealed a phenylalanine tRNA gene whose 3'-end was joined end-to-end with the 5'-end of the 12S rRNA. This analysis first demonstrated the extreme enconomy of genetic material in mammalian mitochondrial DNA.</p

MidExDB: A database of Drosophila CNS midline cell gene expression

<p>Abstract</p> <p>Background</p> <p>The <it>Drosophila </it>CNS midline cells are an excellent model system to study neuronal and glial development because of their diversity of cell types and the relative ease in identifying and studying the function of midline-expressed genes. In situ hybridization experiments generated a large dataset of midline gene expression patterns. To help synthesize these data and make them available to the scientific community, we developed a web-accessible database.</p> <p>Description</p> <p>MidExDB (<it>Drosophila </it>CNS Midline Gene Expression Database) is comprised of images and data from our in situ hybridization experiments that examined midline gene expression. Multiple search tools are available to allow each type of data to be viewed and compared. Descriptions of each midline cell type and their development are included as background information.</p> <p>Conclusion</p> <p>MidExDB integrates large-scale gene expression data with the ability to identify individual cell types providing the foundation for detailed genetic, molecular, and biochemical studies of CNS midline cell neuronal and glial development and function. This information has general relevance for the study of nervous system development in other organisms, and also provides insight into transcriptional regulation.</p

Development of Morphological Diversity of Dendrites in Drosophila by the BTB-Zinc Finger Protein Abrupt

Morphological diversity of dendrites contributes to specialized functions of individual neurons. In the present study, we examined the molecular basis that generates distinct morphological classes of Drosophila dendritic arborization (da) neurons. da neurons are classified into classes I to IV in order of increasing territory size and/or branching complexity. We found that Abrupt (Ab), a BTB-zinc finger protein, is expressed selectively in class I cells. Misexpression of ab in neurons of other classes directed them to take the appearance of cells with smaller and/or less elaborated arbors. Loss of ab functions in class I neurons resulted in malformation of their typical comb-like arbor patterns and generation of supernumerary branch terminals. Together with the results of monitoring dendritic dynamics of ab-misexpressing cells or ab mutant ones, all of the data suggested that Ab endows characteristics of dendritic morphogenesis of the class I neurons

Transcriptional Specificity of Drosophila Dysfusion and the Control of Tracheal Fusion Cell Gene Expression

The Drosophila Dysfusion basic-helix-loop-helix-PAS (bHLH-PAS) protein controls the transcription of genes that mediate tracheal fusion. Dysfusion is highly related to the mammalian Nxf protein that has been implicated in nervous system gene regulation. Toward the goal of understanding how Dysfusion controls fusion cell gene expression, the biochemical properties of Dysfusion were investigated using protein interaction experiments, cell culture-based transcription assays, and in vivo transgenic analyses. Dysfusion dimerizes with the Tango bHLH-PAS protein, and together they act as a DNA binding transcriptional activator. Dysfusion/Tango binds multiple NCGTG binding sites, with the following preference: TCGTG > GCGTG > ACGTG > CCGTG. This binding site promiscuity differs from the restricted binding site preferences of other bHLH-PAS/Tango heterodimers. However, it is identical to the binding site preferences of mammalian Nxf/Arnt, indicating that the specificity is evolutionarily conserved. Germ line transformation experiments using a fragment of the CG13196 Dysfusion target gene allowed identification of a fusion cell enhancer. Experiments in which NCGTG sites were mutated individually and in combination revealed that TCGTG sites were required for fusion cell expression but that the single ACGTG and GCGTG sites present were not. Finally, a reporter transgene containing four tandemly arranged TCGTG elements has strong expression in tracheal fusion cells. Transgenic misexpression of dysfusion further revealed that Dysfusion has the ability to activate transcription in multiple cell types, although it does this most effectively in tracheal cells and can only function at midembryogenesis and later