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Query: UMLS:C0038187 (starvation)
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Gametogenesis in Chlamydomonas reinhardtii has been studied in mating-type plus cells utilizing several different culture conditions, all of which are shown to depend on the depletion of nitrogen from the medium, and the fine structure of gametes prepared under these conditions has been compared by using thin sections of fixed materials. We document alterations in ribosome levels, in chromatin morphology, in starch levels, in the organization of chloroplast membranes, and in the appearance of nuclear envelope and endoplasmic reticulum membranes during gametogenesis. We also noted the acquisition of two new organelles: a mating structure (Friedman, L., A. L. Colwin, and L. H. Colwin. 1968. j. cell Sci. 3:115-128; goodenough, U. W., and R. L. Weiss. 1975. J. Cell Biol. 67:623-637), and Golgi-derived vesicles containing a homogeneous material. We chart the time course of these morphological changes during synchronous gametogenesis. We note that many of these changes may represent adjustments to nitrogen starvation rather than direct features of gametic differentiation, and we also document that cells can differentiate so that they survive conditions of nitrogen starvation for many weeks after they become gametes. We conclude that metabolic alterations, the acquisition of mating ability, and the preparation for long-term survival are all elicited in this organism by nitrogen withdrawal, and we discuss how the various structural alterations observed in this study may relate to these three interrelated avenues of cellular differentiation.
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PMID:Gametic differentiation in Chlamydomonas reinhardtii. I. Production of gametes and their fine structure. 120 15

Two conditional mutants of Chlamydomonas reinhardtii, dif-1 and dif-2, affecting gametic differentiation under conditions of nitrogen (N)-starvation, have been isolated. These mutant cell remain "vegetative" at the restrictive temperature (35 degrees C) in -N medium, as defined by assays of cell body-agglutinin and cell wall lytic enzyme activities in the soluble fractions of cell homogenates. Moreover, the mutants fail to form mating structures at the restrictive temperature, but do so at the permissive temperature (25 degrees C). Temperature-shift experiments show that mutant cells which have differentiated into gametes at 25 degrees C dedifferentiate into "vegetative" cells under N-starvation conditions after transfer to 35 degrees C, but differentiate again into gametes at 25 degrees C. Genetic analyses indicate that the dif-1 and dif-2 genes are recessive and unlinked to each other or to the mating-type locus; the dif-1 phenotype cosegregates with a conditional flagellaless phenotype expressed in both +N and -N medium at the restrictive temperature.
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PMID:Isolation and characterization of Chlamydomonas temperature-sensitive mutants affecting gametic differentiation under nitrogen-starved conditions. 206 64

The unicellular green alga Chlamydomonas reinhardtii responds to sulfate deprivation by producing an arylsulfatase (Lien, T., and O. Schreiner. 1975. Biochim. Biophys. Acta. 384:168-179; Schreiner, O., 1975. Biochim. Biophys. Acta. 384:180-193) and by developing the capacity to transport sulfate more rapidly (our unpublished data). The arylsulfatase activity, detectable 3 h after the transfer of the cells to low sulfate medium (less than or equal to 10 microM sulfate), is a periplasmic protein released into the culture medium by cw15, a cell wall-less mutant of C. reinhardtii. We have purified the derepressible arylsulfatase to homogeneity and have raised monospecific antibodies to it. The protein monomer (67.6 kD) associates into a dimer, and the enzyme activity shows an alkaline pH optimum and a Km of 0.3 mM for p-nitrophenylsulfate. Studies focused on arylsulfatase biosynthesis demonstrate that it is glycosylated and synthesized as a higher molecular mass precursor. The mature protein contains complex N-linked oligosaccharides and the primary translation product has an apparent molecular mass approximately 5 kD larger than the deglycosylated monomer. Since translatable RNA encoding the arylsulfatase can only be detected in cells after sulfate starvation, it is likely that accumulation of the enzyme is regulated at the level of transcription, although posttranscriptional processes may also be involved.
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PMID:Purification and biosynthesis of a derepressible periplasmic arylsulfatase from Chlamydomonas reinhardtii. 333 89

In Chlamydomonas reinhardtii cells, the amount of mRNA coding for the chlorophyll a/b binding proteins of photosystem II (cab II) oscillates in light/dark cycles and in constant dim light. This rhythmic behavior applies to the overall expression of the entire cab II gene family as well as to a single member of the family. The highest mRNA abundance is found in the middle of the subjective day and the lowest in the middle of the subjective night. In constant darkness the cab II mRNA rhythm damps rapidly. The cab II mRNA rhythm persists in non-growing cells under CO2-starvation conditions indicating that the cab II mRNA rhythm is not merely a consequence of cell division, although cell division may influence the amplitude of the cab II mRNA rhythm. The properties of this mRNA oscillation conform to all the major characteristics of circadian rhythms: the period in constant conditions is about 24 h, the rhythm entrains to 24 h light/dark cycles, and the period is temperature compensated. This report is the first demonstration of a circadian rhythm of cab II gene expression in single cells. beta-Tubulin mRNA also shows an oscillation in its abundance in LD cycles and in constant dim light, although its peak-to-trough amplitude is smaller than that of the cab II mRNA rhythm. The beta-tubulin mRNA rhythm peaks in the early night in LD cycles, but in constant illumination, it peaks at about the same circadian phase (i.e., mid-subjective day) as does the cab II mRNA rhythm. Finally, the amount of mRNA coding for mitochondrial cytochrome c is rhythmic in a light/dark cycle but is constant in constant dim light or constant darkness. Surprisingly, this mRNA exhibits a daily oscillation in constant dim light under the specific condition of CO2-depletion.
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PMID:Circadian rhythms of gene expression in Chlamydomonas reinhardtii: circadian cycling of mRNA abundances of cab II, and possibly of beta-tubulin and cytochrome c. 795 2

The non-Mendelian inheritance of chloroplast genes in Chlamydomonas has engaged researchers for decades and has prompted numerous debates regarding molecular mechanisms and evolutionary significance. The hallmarks of chloroplast inheritance in Chlamydomonas are reviewed here, including observations on vegetative haploid cells, somatic hybrids, meiotic zygospores, and vegetative zygotes resulting from sexual reproduction. Models invoked to explain the typical uniparental maternal inheritance of chloroplast genes, and which center upon the presumed existence of sex-specific protectors and destroyers of chloroplast genomes, are briefly discussed. In an effort to bring together the diverse observations on chloroplast gene inheritance in somatic as well as sexual cells, a model is proposed that focuses on organelle DNA turnover as a source of sustenance for the cell during periods of starvation. The salvage/turnover/repair (STOR) model for chloroplast inheritance in Chlamydomonas proposes that as a consequence of the high ploidy of the chloroplast genome, many copies are dispensable; their degradation would provide nucleotides for recombination, repair, RNA synthesis and cell metabolism. The STOR model offers an alternative view of uniparental inheritance as a phenomenon of direct selective benefit to the organism rather than simply being of selfish benefit to the chloroplast genome. These concepts may also have application to other lower eukaryotes that have sexual reproduction coupled with an extended dormancy.
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PMID:The salvage/turnover/repair (STOR) model for uniparental inheritance in Chlamydomonas: DNA as a source of sustenance. 796 52

We have cloned and sequenced the CRY1 gene, encoding ribosomal protein S14 in Chlamydomonas reinhardtii, and found that it is highly similar to S14/rp59 proteins from other organisms, including mammals, Drosophila melanogaster, and Saccharomyces cerevisiae. We isolated a mutant strain resistant to the eukaryotic translational inhibitors cryptopleurine and emetine in which the resistance was due to a missense mutation (CRY1-1) in the CRY1 gene; resistance was dominant in heterozygous stable diploids. Cotransformation experiments using the CRY1-1 gene and the gene for nitrate reductase (NIT1) produced a low level of resistance to cryptopleurine and emetine. Resistance levels were increased when the CRY1-1 gene was placed under the control of a constitutive promoter from the ribulose bisphosphate carboxylase/oxygenase small subunit 2 (RBCS2) gene. We also found that the 5' untranslated region of the CRY1 gene was required for expression of the CRY1-1 transgene. Direct selection of emetine-resistant transformants was possible when transformed cells were first induced to differentiate into gametes by nitrogen starvation and then allowed to dedifferentiate back to vegetative cells before emetine selection was applied. With this transformation protocol, the RBCS2/CRY1-1 dominant selectable marker gene is a powerful tool for many molecular genetic applications in C. reinhardtii.
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PMID:The CRY1 gene in Chlamydomonas reinhardtii: structure and use as a dominant selectable marker for nuclear transformation. 819 40

A promoterless radial spoke protein RSP3 gene has been used to identify promoter regions in the genome of Chlamydomonas reinhardtii. The acceptor strain pf-14 arg7 was transformed with a linearized vector containing the ARG 7.8 gene as a selection marker and a promoterless RSP3 gene. The frequency at which the motility was restored in transformants varied from 2-3%. Several of these were motile only in ammonium-free medium, indicating that the procedure could be used to select inducible promoters. Transformation of nitrogen-starved cells produced about twice as many transformants which were only motile in ammonium-free medium. Since one of the tagging vectors contained an RSP3 gene with a hybridization flag in its 3' untranslated region, it was possible to estimate the size of the new RSP3 transcripts in transformants. The results suggested that in most cases a hybrid RNA was generated consisting of the tagged gene transcript and reporter gene RNA. By 5' RACE, these parts of the new transcripts were amplified and it was shown that the generated DNA fragments could be used to clone a tagged gene. One such example, gene 2BC9, is predicted to code for a mitochondrial matrix protein. The tagging procedure will be optimized for cloning genes induced by nitrogen starvation, the cue for gametogenesis.
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PMID:A promoter trap for Chlamydomonas reinhardtii: development of a gene cloning method using 5' RACE-based probes. 922 72

Chlamydomonas monoica undergoes homothallic sexual reproduction in response to nitrogen starvation. Mating pairs are established in clonal culture via flagellar agglutination and fuse by way of activated mating structures to form the quadriflagellate zygote. The zygote further matures into a dormant diploid zygospore through a series of events that we collectively refer to as zygosporulation. Mutants that arrest development prior to the completion of zygosporulation have been obtained through the use of a variety of mutagens, including ultraviolet irradiation, 5-fluorodeoxyuridine, ethyl methanesulfonate, and methyl methanesulfonate. Complementation analysis indicates that the present mutant collection includes alleles affecting 46 distinct zygote-specific functions. The frequency with which alleles at previously defined loci have been recovered in the most recent mutant searches suggests that as many as 30 additional zygote-specific loci may still remain to be identified. Nevertheless, the present collection should provide a powerful base for ultrastructural, biochemical, and molecular analysis of zygospore morphogenesis and dormancy in Chlamydomonas.
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PMID:The Chlamydomonas zygospore: mutant strains of Chlamydomonas monoica blocked in zygospore morphogenesis comprise 46 complementation groups. 947 27

The light-saturated rate of photosynthetic O2 evolution in Chlamydomonas reinhardtii declined by approximately 75% on a per-cell basis after 4 d of P starvation or 1 d of S starvation. Quantitation of the partial reactions of photosynthetic electron transport demonstrated that the light-saturated rate of photosystem (PS) I activity was unaffected by P or S limitation, whereas light-saturated PSII activity was reduced by more than 50%. This decline in PSII activity correlated with a decline in both the maximal quantum efficiency of PSII and the accumulation of the secondary quinone electron acceptor of PSII nonreducing centers (PSII centers capable of performing a charge separation but unable to reduce the plastoquinone pool). In addition to a decline in the light-saturated rate of O2 evolution, there was reduced efficiency of excitation energy transfer to the reaction centers of PSII (because of dissipation of absorbed light energy as heat and because of a transition to state 2). These findings establish a common suite of alterations in photosynthetic electron transport that results in decreased linear electron flow when C. reinhardtii is limited for either P or S. It was interesting that the decline in the maximum quantum efficiency of PSII and the accumulation of the secondary quinone electron acceptor of PSII nonreducing centers were regulated specifically during S-limited growth by the SacI gene product, which was previously shown to be critical for the acclimation of C. reinhardtii to S limitation (J.P. Davies, F.H. Yildiz, and A.R. Grossman [1996] EMBO J 15: 2150-2159).
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PMID:The regulation of photosynthetic electron transport during nutrient deprivation in Chlamydomonas reinhardtii. 957 82

Gametic differentiation in Chlamydomonas reinhardtii is a two-step process, which is controlled by the sequential action of the two extrinsic signals, nitrogen starvation and blue light. The gamete-specific genes GAS28 and GAS29 are expressed in the late phase of gametogenesis. Their light-induced expression is restricted to cells that have completed the first, nitrogen starvation-activated, phase of differentiation. A comparison of the two genes revealed striking similarities as well as differences. Their most prominent shared feature is an extended sequence homology of over 90% in their 5'-untranslated regions, suggesting a role in translational regulation. GAS28 and GAS29 both encode hydroxyproline-rich proteins (HRGPs) of very similar sizes that exhibit typical features of volvocalean cell wall constituents. GAS28 shows a high degree of homology with the Volvox pherophorin gene family, suggesting a relationship between these genes.
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PMID:Molecular characterization of two light-induced, gamete-specific genes from Chlamydomonas reinhardtii that encode hydroxyproline-rich proteins. 1010 61


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