ABSTRACT
The recent identification of large deposits of sulphates by remote sensing and in situ observations has been considered evidence of the past presence of liquid water on Mars. Here we report the unambiguous detection of diverse phyllosilicates, a family of aqueous alteration products, on the basis of observations by the OMEGA imaging spectrometer on board the Mars Express spacecraft. These minerals are mainly associated with Noachian outcrops, which is consistent with an early active hydrological system, sustaining the long-term contact of igneous minerals with liquid water. We infer that the two main families of hydrated alteration products detected-phyllosilicates and sulphates--result from different formation processes. These occurred during two distinct climatic episodes: an early Noachian Mars, resulting in the formation of hydrated silicates, followed by a more acidic environment, in which sulphates formed.
Subject(s)
Aluminum Silicates/analysis , Aluminum Silicates/chemistry , Climate , Extraterrestrial Environment/chemistry , Mars , Clay , Hydrogen-Ion Concentration , Iron/analysis , Magnesium/analysis , Space Flight , Spacecraft , Sulfates/analysis , Sulfates/chemistry , Water/analysis , Water/chemistryABSTRACT
The multisubunit yeast transcription factor IIIC (TFIIIC) is a multifunctional protein required for promoter recognition, transcription factor IIIB recruitment, and chromatin antirepression. We report the isolation and characterization of TFC7, an essential gene encoding the 55-kDa polypeptide, tau55, present in affinity-purified TFIIIC. tau55 is a chimeric protein generated by an ancient chromosomal rearrangement. Its C-terminal half is essential for cell viability and sufficient to ensure TFIIIC function in DNA binding and transcription assays. The N-terminal half is nonessential and highly similar to a putative yeast protein encoded on another chromosome and to a cyanobacterial protein of unknown function. Partial deletions of the N-terminal domain impaired tau55 function at a high temperature or in media containing glycerol or ethanol, suggesting a link between PolIII transcription and metabolic pathways. Interestingly, tau55 was found, together with TFIIIC subunit tau95, in a protein complex which was distinct from TFIIIC and which may play a role in the regulation of PolIII transcription, possibly in relation to cell metabolism.
Subject(s)
Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/genetics , Transcription Factors, TFIII , Transcription Factors/genetics , Amino Acid Sequence , DNA Polymerase III/genetics , DNA, Fungal/isolation & purification , DNA, Fungal/metabolism , Gene Rearrangement , Molecular Sequence Data , Molecular Weight , Open Reading Frames , Protein Conformation , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/isolation & purification , Saccharomyces cerevisiae/metabolism , Sequence Deletion , Transcription, GeneticABSTRACT
Transcription factor IIIC (TFIIIC) (or tau) is a large multisubunit and multifunctional factor required for transcription of all class III genes in Saccharomyces cerevisiae. It is responsible for promoter recognition and TFIIIB assembly. We report here the cloning and characterization of TFC6, an essential gene encoding the 91-kDa polypeptide, tau91, present in affinity-purified TFIIIC. Tau91 has a predicted molecular mass of 74 kDa. It harbors a central cluster of His and Cys residues and has basic and acidic amino acid regions, but it shows no specific similarity to known proteins or predicted open reading frames. The TFIIIC subunit status of tau91 was established by the following biochemical and genetic evidence. Antibodies to tau91 bound TFIIIC-DNA complexes in gel shift assays; in vivo, a B block-deficient U6 RNA gene (SNR6) harboring GAL4 binding sites was reactivated by fusing the GAL4 DNA binding domain to tau91; and a point mutation in TFC6 (tau91-E330K) was found to suppress the thermosensitive phenotype of a tfc3-G349E mutant affected in the B block binding subunit (tau138). The suppressor mutation alleviated the DNA binding and transcription defects of mutant TFIIIC in vitro. These results indicated that tau91 cooperates with tau138 for DNA binding. Recombinant tau91 by itself did not interact with a tRNA gene, although it showed a strong affinity for single-stranded DNA.
Subject(s)
DNA/metabolism , Saccharomyces cerevisiae/genetics , Transcription Factors, TFIII , Transcription Factors/genetics , Amino Acid Sequence , Binding Sites/genetics , Cloning, Molecular , Molecular Sequence Data , Saccharomyces cerevisiae/metabolism , Sequence Analysis , Transcription Factors/metabolismSubject(s)
Fungal Proteins , RNA Polymerase III , Saccharomyces cerevisiae/metabolism , Transcription Factors , Chemical Precipitation , Chromatography , DNA-Binding Proteins/isolation & purification , DNA-Binding Proteins/metabolism , Fungal Proteins/isolation & purification , Fungal Proteins/metabolism , RNA Polymerase III/isolation & purification , RNA Polymerase III/metabolism , Recombinant Fusion Proteins/metabolism , TATA-Box Binding Protein , Transcription Factor TFIIA , Transcription Factor TFIIB , Transcription Factor TFIIIA , Transcription Factors/classification , Transcription Factors/isolation & purification , Transcription Factors/metabolismABSTRACT
Yeast transcription factor TFIIIB is a multicomponent factor comprised of the TATA-binding protein TBP and of associated factors TFIIIB70 and B". Epitope-tagged or histidine-tagged TFIIIB70 could be quantitatively removed from TFIIIB by affinity chromatography. TBP and B" (apparent mass 160-200 kDa) could be easily separated by gel filtration or ion-exchange chromatography. While only weak interactions were detected between TBP and B", direct binding of [35S]-labeled TBP to membrane-bound TFIIIB70 could be demonstrated in absence of DNA. On tRNA genes, there was no basal level of transcription in the complete absence of TBP. The two characterized TFIIIB components (recombinant rTFIIIB70 and rTBP) and a fraction cochromatographing with B" activity were found to be required for TFIIIC-independent transcription of the TATA-containing U6 RNA gene in vitro. Therefore, beside the TFIIIC-dependent assembly process, each TFIIIB component must have an essential role in DNA binding or RNA polymerase recruitment.
Subject(s)
Fungal Proteins/metabolism , Transcription Factors/metabolism , Chromatography, Affinity , Chromatography, Gel , Chromatography, Ion Exchange , DNA/metabolism , DNA-Binding Proteins/metabolism , Escherichia coli/genetics , Molecular Weight , RNA, Small Nuclear/biosynthesis , Recombinant Proteins/metabolism , TATA-Box Binding Protein , Transcription Factor TFIIIB , Transcription Factors/genetics , Transcription Factors/isolation & purificationABSTRACT
Yeast transcription factor TFIIIB is a multicomponent factor comprised of the TATA-binding protein TBP and of associated factors TFIIIB70 and B". Epitope-tagged or histidine-tagged TFIIIB70 could be quantitatively removed from TFIIIB by affinity chromatography. TBP and B" (apparent mass 160-200 kDa) could be easily separated by gel filtration or ion-exchange chromatography. While only weak interactions were detected between TBP and B", direct binding of [35S]-labeled TBP to membrane-bound TFIIIB70 could be demonstrated in absence of DNA. On tRNA genes, there was no basal level of transcription in the complete absence of TBP. The two characterized TFIIIB components (recombinant rTFIIIB70 and rTBP) and a fraction cochromatographing with B" activity were found to be required for TFIIIC-independent transcription of the TATA-containing U6 RNA gene in vitro. Therefore, beside the TFIIIC-dependent assembly process, each TFIIIB component must have an essential role in DNA binding or RNA polymerase recruitment.
