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A minimal living system and the origin of a protocell.

Based on our current knowledge of molecular biology a living entity is a negentropic system, made of specific catalytic and informational macromolecules, which has the ability to reproduce itself, selectively interact with the environment, increase complexity, undergo mutation and evolve by natural selection. At the cellular level the essential molecular attributes of a minimal living system are considered to be the following: (a) Linear informational molecules, protoRNA (protoDNA) at least 10 mononucleotides long, capable to code for at least 2 amino acids; (b) Code-translating molecules, aminoacyl-proto tRNAs at least 5 mononucleotides long; (c) Protenzymes, oligopeptides of at least 2 amino acids, with measurable catalytic activity for phosphodiester, pyrophosphate and peptide bond formation; (d) Protoribosomes, RNA-peptide complexes which facilitate the interaction of the above three types of molecules, and (e) Protomembranes, liposomes made of amphiphilic lipids and peptides which can provide the semipermeability and the proton gradient necessary for the synthesis of pyrophosphate, ATP and other biochemical compounds. The transition from subcellular Lamarckian evolution to cellular Darwinian evolution required the cooperative interaction, within an internal microenvironment, of the above five types of molecular species.

Biological Evolution↗

Evolution of anticodons.

Anticodons are trinucleotides in transfer RNA (tRNA) molecules. The latter carry amino acids for insertion into the polypeptide sequences of proteins during the translation of messenger RNA (mRNA) molecules. Messenger RNA molecules are transcribed from genes. Evolution of tRNA molecules has resulted in a set of anticodons for the 20 amino acids that are used in protein synthesis. This set of anticodons is slightly different in mitochondrial codes from the set that used in the nuclear "universal" code. Theories for the evolution of the code include frozen accident, doublet expansion, repeating triplets and coevolutionary distribution. The number of codons has always been fixed at 64 by mathematical rules, but because an anticodon may pair with more than one codon, the number of anticodons is only 54 in the universal code, is smaller in mitochondrial codes, and was probably even smaller in archetypal primitive codes. Evidence of anticodon evolution can be seen by comparing mitochondrial codes with the universal code. Codes used by very primitive organisms that are now extinct might have specified fewer amino acids than are now used.

Amino Acids↗

Cell symbiosis [correction of symbioisis] theory: status and implications for the fossil record.

Recent geological treatises have presented three alternative models of the origins of eukaryotes as if they merited equal treatment. However, modern biological techniques, especially nucleic acid and protein sequencing, have clearly established the validity of the symbiotic theory of the origin of eukaryotic organelles. The serial endosymbiotic theory in its most extreme form states that three classes of eukaryotic cell organelles (mitochondria, plastids and undulipodia) originated as free-living bacteria (aerobic respirers, phototrophic bacteria and spirochetes respectively) in association with hosts that become the nucleocytoplasm (Thermoplasma-like archaebacterial hosts). Molecular biological information, primarily derived from ribosomal RNA nucleotide sequencing studies leads to the conclusion that the symbiotic origin theory for both mitochondria and plastids has been proven. The probability of an ancestral archaebacterial-Thermoplasma-like host for the nucleocytoplasm has been rendered more likely by discoveries by Dennis Searcy and his colleagues and Carl Woese and his colleagues. The most equivocal postulate of the symbiotic theory, the origin of undulipodia (cilia and other organelles of motility that develop from kinetosomes is under investigation now. The status of these postulates, as well as their implications for the fossil record, is briefly summarized here.

Bacteria↗

Succession in a microbial mat community: a gaian perspective.

The Gaia hypothesis originally proposed by James E. Lovelock states that the composition, oxidation-reduction state and temperature of the troposphere are actively regulated by the activities of the biota. The gaian concept has been extrapolated to include the composition of surface sediments through the process of biomineralization. The stratified microbial community dominated by the cyanobacterium Microcoleus chthonoplastes is actively involved in the deposition of laminated sediments at Laguna Figueroa, Baja California, Mexico. Unusually heavy rains in the winters of 1979 and 1980 flooded the evaporite flat with up to 3 meters of meteoric water and deposited 5-10 cm of allocthonous sediment. The composition of the microbial community changed as a succession of dominating microbial species ensued, ultimately leading to the recolonization of the surface sediment by the original Microcoleus-dominated community. The resiliency of bacterial communities is suggested to be an important mechanism of gaian control systems.

Cyanobacteria↗

Observational astrochemistry.

Astronomers have identified some 70 molecular species in dense interstellar clouds and the envelopes expelled by evolved stars. Some 75% of these are organic, including molecules with up to 13 atoms and with weights up to 147 daltons. Elements found in interstellar compounds include H, C, N, O, S, Si, and probably Cl. A recent addition to the list of interstellar molecules is the first hydrocarbon ring, cyclopropenylidene (C3H2). Current research in astrochemistry is concentrating on the determination of accurate chemical abundances as a function of environment. Both basic similarities and intriguing differences among clouds are found, which may provide clues to processes associated with cloud evolution and star formation.

Astronomical Phenomena↗

Molecular aspects of adaptation to extreme cold environments.

Here are reviewed and summarized the strategies adopted by living organisms to survive low temperatures, from a molecular and membrane point of view. The presentation is aimed at a wide variety of readers. Two prime examples of connections between biological cold adaptation and the molecular level are (1) antifreeze proteins in fish from cold sea water, (the DNA sequence of the protein gene is now known) (2) the fluidity characteristics of cell membranes in a wide variety of organisms. In model membranes of phospholipids, stabler "s-phases" have recently been found to form at low temperatures. Antarctic endolithic organisms, living just under the surface of rocks, are exposed to long periods of low temperatures, and may develop such phases in their membranes. In the saturated phosphatidyl cholines, only lipids with a restricted range of acyl chain lengths show simultaneously s-phases and a main transition : This restricted range is about the restricted range found in natural membranes. The s-phases also form in the presence of natural cryoprotectants, and may be connected with botanical vernalization.

Acclimatization↗

The Antarctic cold desert and the search for traces of life on Mars.

The cryptoendolithic microorganisms that live inside rocks in the frigid Ross Desert of Antarctica can serve as a terrestrial model for what may have happened to life forms on Mars when the planet became dry and cold. Trace fossils of microbial rock colonization exist in Antarctica, and similar structures could have formed on Mars. In some respects, such trace fossils could be an easier target for life-detection systems than fossils of cellular structures.

Adaptation, Biological↗

The evolution of nucleotides.

There has been much speculation about the types of molecules that were present in the first living forms. Recent discoveries show that RNA is a more versatile molecule than was previously believed, but whether it was ever able, for example, to synthesize its own monomers from available precursors is not yet known. If amino acids coexisted with nucleosides on the prebiotic Earth, then it seems likely that these two classes of molecules would have interacted with each other. We have been studying oligonucleotide-directed peptide bond formation, and during this work we discovered that aminoacylation of the internal 2'-hydroxyl groups of RNA occurred stereoselectively. Investigation of the mechanism of this reaction has been aided by the use of 3'-inosine methyl phosphate (as a simplified model for a dinucleoside monophosphate) and proton nmr spectroscopy of t-butoxycarbonyl-alanyl esters of nucleosides as models for the transition state of the aminoacylation reaction itself.

Acylation↗

Studies on precellular evolution: the encapsulation of polyribonucleotides by liposomes.

Liposomes are 5 to 50 micron vesicles with an internal aqueous environment, whose amphiphilic lipidic components self-assemble into systems with at least one double-layered membrane. Liposomes have been suggested as possible models of precellular systems formed in the early Archean Earth from lipids of non-enzymatic origin. Since it is generally accepted that RNA molecules preceded double-stranded DNA molecules as genetic material, we have studied the encapsulation of polyribonucleotides within liposomes made from dipalmitoyl phosphatidylcholine, and from egg yolk phosphatidylcholine to which cholesterol was added in some cases. The liposomes were prepared under anoxic conditions following the reverse phase evaporation method described by Szoka and Papahadjopoulos. Quantitative determinations show that approximately 50% of the available lipids form liposomes, and that up to 5% of the polyribonucleotides can be entrapped by them. We have also studied the encapsulation of polyribonucleotides in the presence of 1) urea and cyanamide, two non-electrolytes that have been used as prebiotic condensing agents, and 2) of Zn++ and Pb++, two cations employed in the non-enzymatic template-directed synthesis of polyribonucleotides from activated nucleotides.

1,2-Dipalmitoylphosphatidylcholine↗

Comets and life.

Some of the chemical species which have been detected in comets include H2O, HCN, CH3CN, CO, CO2, NH3, CS, C2 and C3. All of these have also been detected in the interstellar medium, indicating a probable relationship between interstellar dust and gas clouds and comets. Laboratory experiments carried out with different mixtures of these molecules give rise to the formation of the biochemical compounds which are necessary for life, such as amino acids, purines, pyrimidines, monosaccharides, etc. However, in spite of suggestions to the contrary, the presence of life in comets is unlikely. On the other hand, the capture of cometary matter by the primitive Earth is considered essential for the development of life on this planet. The amount of cometary carbon-containing matter captured by the Earth, as calculated by different authors, is several times larger than the total amount of organic matter present in the biosphere (10(18)g). The major classes of reactions which were probably involved in the formation of key biochemical compounds are discussed. Our tentative conclusions are that: 1) comets played a predominant role in the emergence of life on our planet, and 2) they are the cosmic connection with extraterrestrial life.

Atmosphere↗

The gamma-irradiation of aqueous hydrogen cyanide in the presence of ferrocyanide or ferricyanide: implications to prebiotic chemistry.

The gamma-irradiation of 0.1 M, O2-free, aqueous HCN was studied in the presence of ferrocyanide or ferricyanide in the concentration range 10(-3) - 10(-5) M. This study was carried out in order to get an insight into the possible role that cyanocomplexes of iron may have played in promoting prebiotic syntheses via the free-radical oligomerization of HCN. It was found that ferrocyanide or ferricyanide have no effect on the irradiation of 0.1 M HCN solutions at concentrations < or = 10(-4) M. At high concentrations, 10(-3) M, they lead to a marked decrease in the conversion of HCN. There was no significant difference due to the oxidation state of iron used, particularly at high doses > or = 100 kGy.

Ammonia↗

A quantitative assay of biologically important compounds in simulated primitive Earth experiments.

A CH4-N2-H2OV gas mixture was subjected to a high voltage (20 kV), high frequency (0.3 MHz) electric discharge. The energy input in the electric discharge was varied from 0.016 to 3.048 MJ mol-1. The chemical yields (G), expressed as the number of molecules formed or destroyed per 100 eV of energy input were calculated for several products. The G values calculated at the lowest energy input were (-CH4) = 6.48; (-N2) = 2.51; (C2H2) = 1.16; (HCN) = 0.215; (CH3CHO) = 0.115; (CH3CH2CHO) = 0.00161; (CH3(CH2)2CHO) = 0.0165; ((CH2CO2H)2) = 0.0000339; (CH4 --> Solid material) = 0.196; (N2 --> Solid material) = 0.00355. This is the first report in prebiotic studies in which the G values of various products in electric discharge experiments are determined. This type of study is needed in order to get a better insight into the relative role of electric discharges on the primitive Earth.

Aldehydes↗

Mineral catalysis of the formation of the phosphodiester bond in aqueous solution: the possible role of montmorillonite clays.

The binding of adenosine to Na(+)-montmorillonite 22A is greater than 5'-AMP, at neutral pH. Adenine derivatives bind more strongly to the clay than the corresponding uracil derivatives. These data are consistent with the protonation of the adenine by the acidic clay surface and a cationic binding of the protonated ring to the anionic clay surface. Other forces must be operative in the binding of uracil derivatives to the clay since the uracil ring system is not basic. The reaction of the 5'-AMP with water soluble carbodiimide in the presence of Na(+)-montmorillonite results in the formation of 2',5'-pApA (18.9%), 3',5'-pApA (11%), and AppA (4.8%). When poly(U) is used in place of the clay the product yields are 2',5'-pApA (15.5%), 3',5'-pApA (3.7%) and AppA (14.9%). The cyclic nucleotide, c(pA)2 is also formed when poly(U) is used. AppA is the principal reaction product when neither clay nor poly(U) is present in the reaction mixture. When 2'-deoxy-5'-AMP reacts with carbodiimide in the presence of Na(+)-montmorillonite 22A the products are dpApA (4.8%), dAppApA (4.5%) and dAppA (17.4%). Cyclic 3',5'-dAMP is the main product (14%) of the reaction of 2'-deoxy-3'-AMP.

Adenine↗

Nucleic acid analogues and the origins of replication.

Recent interest in the properties of "nucleic acid-like structures" has been stimulated by difficulties encountered in the synthesis and nonenzymatic oligomerization of nucleotides. However, none of the newly proposed monomers has yet been synthesized in a plausibly prebiotic manner. Arguments are presented that analogues based on 8-hydroxymethyladenine and 5-hydroxymethyluracil are promising candidates for primitive nucleotide precursors.

Adenine↗

Early martian environments: the Antarctic and other terrestrial analogs.

The comparability of the early environments of Mars and Earth, and the biological evolution which occurred on early earth, motivates serious consideration of the possibility of an early martian biota. Environments which could have contained this early martian life and which may presently contain evidence of this former life include aquatic, ice, soil, and rock habitats. Several analogs of these potential early martian environments, which can provide useful information in searching for extinct life on Mars, are currently available for study on Earth. These terrestrial analogs include the perennially ice-covered lakes and sandstone rocks in the Polar Deserts of Antarctica, surface of snowfields and glaciers, desert soils, geothermal springs, and deep subsurface environments.

Antarctic Regions↗

Life on Mars: how it disappeared (if it was ever there).

The cryptoendolithic microbial community in the Ross Desert (McMurdo Dry Valleys) of Antarctica exists at temperatures significantly below the temperature optima of the primary producers. Surviving near the limit of their physiological adaptability, the organisms are under severe environmental stress, so further deterioration in the environment results in cell damage and death. The sequence of events leading to extinction is considered to be a terrestrial analog for disappearance of possible life on early Mars. Progressive stages of cell damage and death in the Ross Desert material are documented with transmission electron microscopy.

Antarctic Regions↗

Planetary protection issues for sample return missions.

Sample return missions from a comet nucleus and the Mars surface are currently under study in the US, USSR, and by ESA. Guidance on Planetary Protection (PP) issues is needed by mission scientists and engineers for incorporation into various elements of mission design studies. Although COSPAR has promulgated international policy on PP for various classes of solar system exploration missions, the applicability of this policy to sample return missions, in particular, remains vague. In this paper, we propose a set of implementing procedures to maintain the scientific integrity of these samples. We also propose that these same procedures will automatically assure that COSPAR-derived PP guidelines are achieved. The recommendations discussed here are the first step toward development of official COSPAR implementation requirements for sample return missions.

Containment of Biohazards↗

Observational astrochemistry: recent results.

More than 80 molecular species have now been observed by astronomers in the dense interstellar clouds where stars and planets form or in the envelopes expelled by evolved stars. Elemental constituents of these compounds include all of the "biogenic" elements, hydrogen, carbon, nitrogen, oxygen, sulfur, and (most recently) phosphorus. In addition, silicon is found in several molecules, and a series of metal halides have recently been detected in the outflowing envelope of a nearby carbon star. Additions to the list of known interstellar molecules since the last COSPAR meeting are discussed individually. Recent measurements of the hydrogen isotopic fractionation for the cyclic molecule C3H2 are described; values up to 10,000 times the cosmic deuterium-to-hydrogen ratio are found. Knowledge of the chemical reservoirs for the major volatile elements and a comparison between observed molecular abundances and theoretical models are both discussed.

Acetone↗