Nucleotide sequence of the tcml gene (ribosomal protein L3) of Saccharomyces cerevisiae

Schultz, Loren D.; Friesen, James D.

doi:10.1128/jb.155.1.8-14.1983

Cited by 122 publications

(20 citation statements)

References 38 publications

(40 reference statements)

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“…Our results also reveal that it is possible to use trichodermin resistance as a dominant selection marker for transformation of C. aibicans, gene tcm1 codes for a S. cerevisiae modified ribosomal protein, L3, of the 60S subunit (Schultz and Friesen, 1983) and it seems to be expressed in C. aibicans, thus leading to resistant transformant clones. The use of dominant selection markers represents a clear advantage from the point of view of the development of vectors, since it may enable strategies of genetic transformation that will not depend on the use of strains with specific markers.…”

Section: Deveiopment Of Candida Transforming Plasmids With a Dominantmentioning

confidence: 69%

A reorganized Candida albicans DNA sequence promoting homologous non‐integrative genetic transformation

Herreros

García-Sáez

Nombela

et al. 1992

Molecular Microbiology

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In order to develop plasmids adequate for non-integrative genetic transformation of Candida albicans, a DNA fragment of 15.3 kb was cloned from this organism on the basis of its capacity to convert the integrative Saccharomyces cerevisiae vector YIp5 into a non-integrative one. Southern hybridization analysis, carried out with a labelled DNA probe of 3.6 kb derived from the cloned fragment, showed that it consisted of C. albicans DNA, the hybridization pattern indicating that the corresponding sequences were homologous to several chromosomal regions. The size of the C. albicans DNA promoting autonomous replication in S. cerevisiae was substantially reduced by subcloning. A 5.1 kb subfragment, defined by BamHI and SalI restriction sites, retained autonomous replication sequences (ARS) functional in the heterologous S. cerevisiae system and in C. albicans, when inserted in plasmid constructions that carried a S. cerevisiae trichodermin-resistance gene (tcm1) as selection marker. C. albicans transformants were both of the integrative and the non-integrative type and the plasmids recovered from the latter very often carried a reorganized ARS, indicating that recombination of the inserted ARS DNA had occurred in the homologous host. Successive reorganizations of the ARS insert in C. albicans eventually led to a more stable and much smaller fragment of 687 bp that was subsequently recovered unchanged from transformants. Sequence analysis of the 687 bp fragment revealed four 11-base blocks, rich in A+T, that carried the essential consensus sequence considered relevant for yeast ARS elements in addition to other features also described as characteristic of yeast replication origins.

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Section: Deveiopment Of Candida Transforming Plasmids With a Dominantmentioning

confidence: 69%

A reorganized Candida albicans DNA sequence promoting homologous non‐integrative genetic transformation

Herreros

García-Sáez

Nombela

et al. 1992

Molecular Microbiology

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“…Mutants of S. cerevisiae carrying the leu2 mutation in addition to the temperature-sensitive mutation rna2, rna4, or rnall (strains YR02-92, YRO4-304, YR11-182, respectively; Table 1) were transformed with one of two libraries of DNA fragments prepared from S. cerevisiae. One of these libraries (from which RNA2+ was isolated) was from a total BamHI digest of strain CLP1 DNA (13) carried on pJZ1 (47), and the other library (from which RNA4+ and RNAJJ+ were isolated) was a partial HindIII digestion of strain S288C DNA carried on pYF91 (51) (the cloning vectors and typical shuttle vectors containing a 2 ,um plasmid replication origin, the LEU2+ S. cerevisiae gene, and pBR322 sequences). Transformants which were both Leu+ and temperature resistant when grown at 37°C were selected.…”

Section: Resultsmentioning

confidence: 99%

“…E. coli transformation was performed by the method of Mandel and Higa (33). The shuttle vectors used for DNA cloning were pYF91 (51) or pJZ1 (47). pJH18 (supplied by Jim Haber) is a pBR322 derivative containing a 1.1-kilobase Hindlll DNA fragment containing the URA3+ gene.…”

Section: Methodsmentioning

confidence: 99%

Isolation and characterization of the RNA2+, RNA4+, and RNA11+ genes of Saccharomyces cerevisiae

Soltyk¹,

Tropak²,

Friesen³

1984

J Bacteriol

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We used genetic complementation to isolate DNA fragments that encode the Saccharomyces cerevisiae genes RNA2+, RNA4+, and RNA I+ and to localize the genes on the cloned DNA fragments. RNA blot-hybridization analyses coupled with genetic analyses indicated that RNA2+ is coded by a 3.0-kilobase (kb) transcript, RNA4+ is coded by a 1.6-kb transcript, and RNAH + is coded by a 1.3-kb or a 1.7-kb transcript or both; none of the cloned genes contains detectable introns. All three genes were transcribed into messages of very low abundance (-20 times lower than a ribosomal protein message). DNA blot-hybridization revealed that all cloned genes are represented only once in the yeast chromosome. mRNA for RNA2+ and RNA4+ is produced in approximate proportion to gene dosage, whereas RNA1l+ transcription appears to be not nearly so dependent on gene dosage. On a medium-copy plasmid (5 to 10 copies per cell), each cloned gene complemented mutations only in its own gene, indicating that each gene encodes a unique function. Genetic analysis by integrative transformation indicated that we cloned the RNA2+, RNA4+, and RNA1ll structural genes and not second-site suppressors.on July 16, 2020 by guest

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“…The deduced amino acid sequence showed significant homology to tcm1, which encodes RpL3 of S. cerevisiae . It is interesting to note that tcm1 has been implicated in resistance of S. cerevisiae to trichodermin (an inhibitor of ribosome peptidyl transferase activity) [26,27]. Although many strains of Afu have been shown to be resistant to antibiotics, only limited information is yet available regarding the genes conferring antibiotic resistance to Afu .…”

Section: Discussionmentioning

confidence: 99%

cDNA cloning, expression and characterization of an allergenic L3 ribosomal protein of Aspergillus fumigatus

Saxena

Madan

Muralidhar

et al. 2003

Clinical and Experimental Immunology

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SUMMARYAspergillus fumigatus ( Afu ) is an important fungal pathogen causing allergic and invasive respiratory disorders. A plethora of multi-functional allergens/antigens secreted by Afu have been implicated in pathogenesis. The present study was undertaken to identify and characterize novel Afu allergen/antigen by cDNA library approach. cDNA library of Afu was immunoscreened with pooled sera of allergic bronchopulmonary aspergillosis (ABPA) patients. The cDNA clone, TS1, reacting significantly with specific IgG antibodies, was selected. cDNA was subcloned and expressed in Escherichia coli. Sequencing of the cDNA revealed an open reading frame (ORF) of 1179 bases coding for a protein with an approximate molecular weight of 44 kDa. Immunoreactivity of the recombinant TS1 protein (rTS1) was evaluated by ELISA and Western blot analysis using pooled sera of ABPA patients. The rTS1 exhibited binding to specific IgG and IgE antibodies present in sera of ABPA patients. The deduced amino acid sequence showed homology to 60S ribosomal protein L3 (RpL3) of Aspergillus nidulans , Saccharomyces cerevisiae and Homo sapiens . The RpL3 of S. cerevesiae, tcm1 , to which TS1 sequence shows significant homology (72% identity), is known to be responsible for conferring resistance against trichodermin (antibiotic, inhibiting protein synthesis). The present study has led to identification, cloning and expression of a 44-kDa novel allergen/antigen of Afu with sequence homology to L3 ribosomal protein with a probable role in resistance of Afu to antifungal drugs. Sixty-four per cent sequence identity of Afu RpL3 with human RpL3 and common regions in their predicted epitopes suggest a possibility of involvement of Afu RpL3 in autoimmune reactions due to molecular mimicry.

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Nucleotide sequence of the tcml gene (ribosomal protein L3) of Saccharomyces cerevisiae

Cited by 122 publications

References 38 publications

A reorganized Candida albicans DNA sequence promoting homologous non‐integrative genetic transformation

A reorganized Candida albicans DNA sequence promoting homologous non‐integrative genetic transformation

Isolation and characterization of the RNA2+, RNA4+, and RNA11+ genes of Saccharomyces cerevisiae

cDNA cloning, expression and characterization of an allergenic L3 ribosomal protein of Aspergillus fumigatus

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