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RNA oxidative damage and ribosomal RNA surveillance under oxidative stress
Title
RNA oxidative damage and ribosomal RNA surveillance under oxidative stress.
Creator
Liu, Min, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description
We have studies oxidative damage of RNA, a major type of cellular macromolecules. RNA is a primary target of reactive oxygen species (ROS). Under oxidative stress, most nucleic acid damages in Escherichia coli (E.coli) are present in RNA as shown by high levels of 8-oxo-G, an oxidized form of guanine. Increased RNA oxidation is closely correlated to cell death under oxidative stress. Surprisingly, neither RNA structure nor association with proteins protects RNA from oxidation... Our results...
Show moreWe have studies oxidative damage of RNA, a major type of cellular macromolecules. RNA is a primary target of reactive oxygen species (ROS). Under oxidative stress, most nucleic acid damages in Escherichia coli (E.coli) are present in RNA as shown by high levels of 8-oxo-G, an oxidized form of guanine. Increased RNA oxidation is closely correlated to cell death under oxidative stress. Surprisingly, neither RNA structure nor association with proteins protects RNA from oxidation... Our results demonstrate a major role for RNA degradation in controlling oxidized RNA. We have identified activities that may work in specific pathways for selectively degrading damaged RNA. These activities may play pivotal rold in controlling oxidized RNA and protecting cells under oxidative stress.
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Date Issued
2012
PURL
http://purl.flvc.org/FAU/3355620
Subject Headings
RNA, Metabolism, Cellular signal transduction, Genetic translation, Molecular biology
Format
Document (PDF)
The Application of Yeast Three-Hybrid Technology in the Mexican Axolotl (Ambystoma mexicanum) System.
Title
The Application of Yeast Three-Hybrid Technology in the Mexican Axolotl (Ambystoma mexicanum) System.
Creator
Maier, Jennifer A., Lemanski, Larry F., Florida Atlantic University
Abstract/Description
The Mexican axolotl, Ambystoma mexicanum, possesses a naturally-occurring lethal mutation, designated gene "c", for cardiac non-function. Hearts form but fail to beat, lack organized myofibrils, and are deficient in tropomyosin. Treatment with a noncoding RNA MIR (Myofibril-Inducing RNA) rescues this defect in organ culture. Rescued mutant hearts have restored tropomyosin, form organized myofibrils, and beat vigorously. Studies to elucidate the mechanism of MIR heart rescue are underway....
Show moreThe Mexican axolotl, Ambystoma mexicanum, possesses a naturally-occurring lethal mutation, designated gene "c", for cardiac non-function. Hearts form but fail to beat, lack organized myofibrils, and are deficient in tropomyosin. Treatment with a noncoding RNA MIR (Myofibril-Inducing RNA) rescues this defect in organ culture. Rescued mutant hearts have restored tropomyosin, form organized myofibrils, and beat vigorously. Studies to elucidate the mechanism of MIR heart rescue are underway. Current evidence suggests that MIR acts by binding with at least two proteins. The yeast three-hybrid system is being used to screen an axolotl eDNA library for these two proteins and other possible MIR-binding candidates. This is a method utilizing two hybrid proteins and a hybrid RNA. An interaction between these three components will activate the expression of reporter genes, whose activity is assayed through phenotypical and biochemical methods. In this study, the protocol for yeast three-hybrid technology is being established for analyzing the MIR in the Mexican axolotl, cardiac mutant animal model.
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Date Issued
2007
PURL
http://purl.flvc.org/fau/fd/FA00000793
Subject Headings
Axolotls--Development, Heart--Hypertrophy, Genetic translation, Molecular genetics--Research
Format
Document (PDF)
Metalloprotease Profiling in Melanoma
Title
Metalloprotease Profiling in Melanoma.
Creator
Giricz, Orsolya, Florida Atlantic University, Fields, Gregg B., Charles E. Schmidt College of Science, Department of Chemistry and Biochemistry
Abstract/Description
The proteolytic activities of the ADAM (a disintegrin and metalloproteinase), ADAMTS (a disintegrin and metalloproteinases with thrombospondin motifs) and MMP (matrix metalloproteinase) protein families play important roles in normal and multiple pathological conditions. These metalloproteases have potential implications in the degradation of the extracellular matrix and in the processing of bioactive molecules. Under pathological conditions these proteases are involved in many diverse...
Show moreThe proteolytic activities of the ADAM (a disintegrin and metalloproteinase), ADAMTS (a disintegrin and metalloproteinases with thrombospondin motifs) and MMP (matrix metalloproteinase) protein families play important roles in normal and multiple pathological conditions. These metalloproteases have potential implications in the degradation of the extracellular matrix and in the processing of bioactive molecules. Under pathological conditions these proteases are involved in many diverse processes from tumor cell migration to cartilage destruction in rheumatoid arthritis. In the present study, the gene expression levels of six ADAMs, eight MMPs, and four ADAMTSs were analyzed by Real Time PCR. RNA was isolated from multiple normal fibroblast and metastatic melanoma cell lines, as well as the isogenic normal tissue and tumor samples. This method allowed for detected changes in mRNA expressiOn of the individual metalloproteainase genes to be compared between normal and metastatic states, and also between tissue and cultured cells. Substantial differences have been observed in the level of ADAM and MMP mRNA expression between tissue and cell lines. In general, the level of expression is several folds higher in cultured cells compared to the isogenic tissue they are derived from. Protein microarrays were utilized in order to evaluate the correlations between MMP and TIMP mRNA copy numbers and protein abundance in cell culture. In several cases, distinct differences were observed regarding the localization of the proteins examined. In order to determine if the metalloprotease genes that were elevated at the level of RNA expression produce functional proteins, the foundations of an in situ FRET assay have been established. This will greatly aid in a better understanding of the behavior of metallopeptidases in a cellular context.
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Date Issued
2008
PURL
http://purl.flvc.org/fau/fd/FA00000859
Subject Headings
Tumor markers--Research, Metalloproteinases--Inhibitors, Melanoma--Research, Proteins--Synthesis, Genetic translation
Format
Document (PDF)