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At least 487 records · Page 27

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NASA NTRS1982

The chemical structure of DNA sequence signals for RNA transcription

The proposed recognition sites for RNA transcription for E. coli NRA polymerase, bacteriophage T7 RNA polymerase, and eukaryotic RNA polymerase Pol II are evaluated in the light of the requirements for efficient recognition. It is shown that although there is good experimental evidence that specific nucleic acid sequence patterns are involved in transcriptional regulation in bacteria and bacterial viruses, among the sequences now available, only in the case of the promoters recognized by bacteriophage T7 polymerase does it seem likely that the pattern is sufficient. It is concluded that the eukaryotic pattern that is investigated is not restrictive enough to serve as a recognition site.

George, D. G.↗

NASA NTRS1983

Investigation of compounds essential for the origin of life

Nucleic acid sequencing as a technique to determine the chemical and biological evolution of certain prokaryotic metabolic pathways is discussed. Protein in data and a microbiological organization of the prokaryotes is included.

Dayhoff, M. O.↗

NASA NTRS1983

Sequence evidence for the symbiotic origins of chloroplasts and mitochondria

The origin of mitochondria and chloroplasts is investigated on the basis of prokaryotic and early-eukaryotic evolutionary trees derived from protein and nucleic-acid sequences by the method of Dayhoff (1979). Trees for bacterial ferrodoxins, 5S ribosomal RNA, c-type cytochromes, the lipid-binding subunit of ATPase, and dihydrofolate reductase are presented and discussed. Good agreement among the trees is found, and it is argued that the mitochondria and chloroplasts evolved by multiple symbiotic events.

George, D. G.↗

NASA NTRS1984

On the stereoselective aminoacylation of RNA

Gabbay and Kleinman (1970) have found that stereospecific complex formation (noncovalent) occurs between nucleic acids and a number of derivatives of amino acids. However, until recently, chiral selection in any nonenzymatic RNA-aminoacylation reaction was unknown. Profy and Usher (1984) reported that aminoacylation of the 'internal' 2-prime-ester occurred with a significant amount of stereoselection. Profy and Usher (1984) have also observed that aminoacylation of the 'internal' 2-prime-hydroxyl groups of polyribonucleotides by the imidazolide of N-3,5-dinitrobenzoylalanine occurs with chiral selection. In order to obtain further information regarding the considered phenomena, a systematic investigation was initiated of the factors which contribute to the observed stereoselectivity of the aminoacylation reaction. In the present paper, the effect of a change in the amino acid from alanine to leucine is considered along with an investigation of the D- and L-alanyl internal' 2-prime esters of the dinucleoside monophosphate of 3-prime,5-prime-ApA.

Usher, D. A.↗

NASA NTRS1985

Utilization of non-conventional systems for conversion of biomass to food components: Potential for utilization of algae in engineered foods

The major nutritional components of the green algae (Scenedesmus obliquus) grown in a Constant Cell density Apparatus were determined. Suitable methodology to prepare proteins from which three major undesirable components of these cells (i.e., cell walls, nucleic acids, and pigments) were either removed or substantially reduced was developed. Results showed that processing of green algae to protein isolate enhances its potential nutritional and organoleptic acceptability as a diet component in a Controlled Ecological Life Support System.

Karel, M.↗

NASA NTRS1986

Potential for utilization of algal biomass for components of the diet in CELSS

The major nutritional components of the green algae (Scenedesmus obliquus) grown in a Constant Cell Density Apparatus were determined. Suitable methodology to prepare proteins from which three major undesirable components of these cells (i.e., cell walls, nucleic acids, and pigments) were either removed or substantially reduced was developed. Results showed that processing of green algae to protein isolate enhances is potential nutritional and organoleptic acceptability as a diet component in controlled Ecological Life Support System.

Kamarei, A. R.↗

NASA NTRS1986

Synthesis and analysis in studies of chemical evolution

Studies of the various processes that may have given rise to life on the Earth have demonstrated the appropriateness of an approach that makes use of analysis and synthesis. Analysis of extraterrestrial samples in the form of meteorites has demonstrated the presence of several precursors of biomolecules, most notably a full suite of nucleic acid bases and nucleotides of biological significance. These species were determined after exhaustive extraction of the sample and subsequent analysis using HPLC, GC, MS, and GC-MS. Procedural blanks indicate that these molecules are likely not the result of contamination during the extraction and analysis process. Similar species were found as products of spark discharge experiments in atmospheres thought to mimic primitive Earth conditions. These results indicate that the basic chemistry underlying these syntheses is common, and that life may not be unique to the Earth. Studies underway in the laboratory make use of proton nuclear magnetic resonance spectroscopy as a probe to assess associations between selected amino acids and any of several nucleotides comprising their genetic code and genetic anticode sequences. These studies demonstrate a clear selectivity by the anticode sequences, thus confirming the hydrophobicity studies performed by Lacey et al. These studies further support the contention that life is likely a natural result of the physics and chemistry of the universe.

Ponnamperuma, C.↗

NASA NTRS1986

Stereoselective aminoacylation of RNA

Prebiotic chemistry is faced with a major problem: how could a controlled and selective reaction occur, when there is present in the same solution a large number of alternative possible coreactants? This problem is solved in the modern cell by the presence of enzymes, which are not only highly efficient and controllable catalysts, but which also can impose on their substrates a precise structural requirement. However, enzymes are the result of billions of years of evolution, and we cannot invoke them as prebiotic catalysts. One approach to solving this problem in the prebiotic context is to make use of template-directed reactions. These reactions increase the number of structural requirements that must be simultaneously present in a molecule for it to be able to react, and thereby increase the selectivity of the reaction. They also can give a large increase in the rate of a reaction, if the template constrains two potential coreactants to lie close together. A third benefit is that information that is present in the template molecule can be passed on to the product molecules. If the earliest organisms were based on proteins and nucleic acids, then the investigation of peptide synthesis on an oligonucleotide template is highly relevant to the study of the origin of life.

Usher, D. A.↗

NASA NTRS1986

Protein crystal growth; Proceedings of the First International Conference, Stanford University, CA, August 14-16, 1985

Papers are presented on mechanisms of nucleation and growth of protein crystals, the role of purification in the crystallization of proteins and nucleic acids, and the effect of chemical impurities in polyethylene glycol on macromolecular crystallization. Also considered are growth kinetics of tetragonal lysozyme crystals, thermodynamic and kinetic considerations for crystal growth of complex molecules from solution, protein single-crystal growth under microgravity, and growth of organic crystals in a microgravity environment. Papers are also presented on preliminary investigations of protein crystal growth using the Space Shuttle, convective diffusion in protein crystal growth, and the growth and characterization of membrane protein crystals.

Feigelson, R. S.↗

NASA NTRS1989

Utilization of non-conventional systems for conversion of biomass to food components

The potential use of micro-algae in yielding useful macronutrients for the CELSS is investigated. Algal proteins were isolated and characterized from green algae (Scenedesmus obliquus) grown under controlled conditions. The RNA and DNA contents were determined, and methodology for reduction of the nucleic acid content to acceptable levels developed. Lipid extraction procedures using supercritical fluids were tailored to removal of undesirable lipids and pigments. Initial steps toward preparation of model foods for potential use in the CELSS were taken. The goal was to fabricate food products which contain isolated algal macronutrients such as proteins and lipids and also some components derived from higher plants including wheat flour, soy flour, potato powder (flakes), soy oil, and corn syrup.

Karel, M.↗

NASA NTRS1989

Model of early self-replication based on covalent complementarity for a copolymer of glycerate-3-phosphate and glycerol-3-phosphate

Glyceraldehyde-3-phosphate acts as the substrate in a model of early self-replication of a phosphodiester copolymer of glycerate-3-phosphate and glycerol-3-phosphate. This model of self-replication is based on covalent complementarity in which information transfer is mediated by a single covalent bond, in contrast to multiple weak interactions that establish complementarity in nucleic acid replication. This replication model is connected to contemporary biochemistry through its use of glyceraldehyde-3-phosphate, a central metabolite of glycolysis and photosynthesis.

Weber, Arthur L.↗

NASA NTRS1989

Utilization of non-conventional systems for conversion of biomass to food components

Described here is work accomplished in investigating the potential use of micro-algae in yielding useful macronutrients for closed ecological life support systems in space habitats. Analysis of the chemical composition of the blue-green alga Synechoccus 6311 was done in the present work, and was compared to values found in previous work on the green algae Scenedesmus obliquus. Similar values were obtained for proteins, and lower values for nucleic acids and lipids. A second part of the work involved fabrication of food products containing various levels of incorporated algae (S. obliquus) proteins and/or lipids. Protein isolate was incorporated into a variety of food products such as bran muffins, fettuccine (spinach noodle imitation), and chocolate chip cookies. In the sensory analysis, the greenish color of the bran muffins and cookies was not found to be objectionable. The mild spinachy flavor was less detectable in chocolate chip cookies than in bran muffins. The color and taste of the algae noodles were found to be pleasant and compared well with commercially available spinach noodles.

Karel, M.↗

NASA NTRS1989

Model of early self-replication based on covalent complementarity for a copolymer of glycerate-3-phosphate and glycerol-3-phosphate

Weber, Arthur L.↗

NASA NTRS1991

Molecular bases for unity and diversity in organic evolution

The origin of biological information has been ascribed at various times to DNA, RNA, or protein. The origin of nucleic acids without the action of prior informed protein has not been supported by plausible experiments, although such possibilities have been examined. The behavior of thermal proteins and of the microspheres selfassembled therefrom explain the origin of the first cells, the first membrane, the first reproduction cycle, ancient metabolism including ATP-aided synthesis of peptides and polynucleotides, growth, bioelectricity, and polybiofunctionality in general.

Fox, S. W.↗

NASA NTRS1992

Possible prebiotic significance of polyamines in the condensation, protection, encapsulation, and biological properties of DNA

While DNA which has undergone ionic condensation with Co(3+)(NH3)6 is resistant to the action of the endonuclase DNAse I, in much the same way as DNA condensed with spermidine, it was significantly less active in transcription with the E. coli RNA polymerase than DNA-spermidine condensed forms. Although both compacted forms of DNA were more efficiently encapsulated into neutral liposomes, negatively charged liposomes were seldom formed in the presence of the present, positive ion-condensed DNA; spermidine is accordingly proposed as a plausible prebiotic DNA-condensing agent. Attention is given to the relevance of the polyimide-nucleic acids complexes in the evolution of life.

Baeza, Isabel↗

NASA NTRS1996

Separation of 'Uncharged' Oligodeoxynucleotide Analogs by Anion-Exchange Chromatography at High pH

Ion-exchange chromatography is a well-established method for the analysis and purification of phosphodiester-linked oligonucleotides. If elution is carried out under alkaline conditions, the secondary structure of G- and C-rich oligomers is disrupted. Furthermore, elution times become more sensitive to the G and T content of the oligomer, because G and T are deprotonated at pH 10. In recent work on peptide-nucleic acids (PNAs) we noted that mixtures of PNA oligomers G(sub 4), G(sub 6), G(sub 8), and G9(sub 10) are readily separated by elution at pH 12 on an RPC-5 column. Here we show that this separation method is more generally applicable.

Schmidt, Jurgen G.↗

NASA NTRS1995

The Relative Reactivity of Deoxyribose and Ribose: Did DNA Come Before RNA?

If it is assumed that there was a precursor to the ribose-phosphate backbone of RNA in the preRNA world (such as peptide nucleic acid), then the entry of various sugars into the genetic material may be related to the stability and non-enzymatic reactivity of the aldose. The rate of decomposition of 2-deoxyribose has been determined to be 1/3 that of ribose. In addition we have measured the amount of free aldehyde by H-1 and C-13 NMR and find that it has approximately 0.15% free aldehyde compared to 0.05% for ribose at 25 C. This suggests that deoxyribose would be significantly more reactive with early bases in the absence of enzymes. This is confirmed by urazole and deoxyribose reacting to form the deoxynucleoside 45 times faster as 25 C than urazole reacts with ribose to form the Ribonucleoside. Urazole is a potential precursor of uracil and is a plausible prebiotic compound which reacts with aldoses to form nucleosides. Thus the non-enzymatic reactivity of deoxyribose would favor its early use over ribose until enzymes could change the relative reactivities. Most of the reasons that RNA is presumed to have come before DNA are extrapolations back from contemporary metabolism (e.g. the abundance of ribose based coenzymes, the biosynthesis of histidine, deoxyribonucleotides are synthesized from ribonucleotides, etc.). It is very difficult to reconstruct biochemical pathways much before the last common ancestor, and it is even more difficult to do more than guess at the biochemistry of very early self-replicating systems. Thus we believe that these reasons are not compelling and that the non-enzymatic chemistry may be more important than enzymatic pathways for constructing the earliest of biochemical pathways. While the RNA world has been discussed at great length, there has not been an exploration of the transition out of the RNA world. We have constructed many possible schemes of genetic takeover events from preRNA to modern DNA, RNA, protein system which could generate the RNA metabolic fossils we see today.

Dworkin, Jason P.↗

NASA NTRS1998

An Advanced Approach to Simultaneous Monitoring of Multiple Bacteria in Space

This interim report describes progress on the development of spacecraft-compatible methods of nucleic acid preparation and cleanup. Grantee found that performance of a widely-known protocol could be greatly enhanced by the addition of a sizing step to eliminate acids. Testing of alternative adsorptive methods is underway. Grantee also confirmed and extended observation of precipitation of DNA by condensing agents such a spermine at low ionic strength.

Eggers, Mitch↗