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Interactions between chromosomal omnipotent suppressors and extrachromosomal effectors in Saccharomyces cerevisiae.

Chromosomal omnipotent suppressor mutations recovered in psi+ strains of Saccharomyces cerevisiae were brought into psi- cytoplasm. SUP46, SUP138 and SUP139 acted as dominant omnipotent suppressors in the psi- cytoplasm though their suppressor activity was substantially reduced. SUP46 and SUP138 conferred recessive thermosensitivity and antibiotic sensitivity in psi- cytoplasm as in psi+ cytoplasm. On the other hand, sup111 through sup115, which acted as recessive omnipotent suppressors in the psi+ cytoplasm, manifested no, or very low, suppressor activity in the psi- cytoplasm. They, however, still enhanced the efficiency of the SUP29 tRNA suppressor in psi- cytoplasm. A multicopy plasmid carrying the wild-type SUP35 gene enhanced the efficiency of sup111 in psi- cytoplasm.

Extrachromosomal Inheritance↗

rps10, unreported for plastid DNAs, is located on the cyanelle genome of Cyanophora paradoxa and is cotranscribed with the str operon genes.

rps10, encoding the plastid ribosomal protein S10, is a nuclear gene in higher plants and green algae, and is missing from the large ribosomal protein gene cluster of chlorophyll b-type plastids that contains components of the prokaryotic S10, spc and alpha operons. The cyanelle genome of Cyanophora paradoxa is shown to harbor rps10 as another specific feature of its organization. However, this novel plastid gene is not contiguous with the genes of the "S10" operon, but is adjacent to, and cotranscribed with, the str operon, a trait also found in archaebacteria.

Amino Acid Sequence↗

Expression of CMS-unique and flanking mitochondrial DNA sequences in Phaseolus vulgaris L.

The expression of mitochondrial DNA sequences unique to a cytoplasmically male-sterile (CMS) line of Phaseolus vulgaris was investigated. RNA-blot hybridizations with strand-specific probes demonstrated CMS-unique transcripts (7.0, 6.8, 4.7, 3.3 and 2.8 kb) to be in the sense orientation with respect to the longest open reading frames within the CMS-unique region. Hybridizations revealed co-transcription of CMS-unique and upstream, atpA-coding sequences to generate the 6.8-kb RNA. However, hybridizations with CMS-unique and flanking DNA probes accounted for only 4.9 kb of the longest and most abundant (7.0 kb) CMS-unique transcript, providing indirect evidence for the involvement of a splicing process in the generation of this transcript. Sedimentation experiments demonstrated the association of 7.0- and 6.8-kb CMS-unique transcripts with polyribosomes in seedlings and floral buds of a CMS line and a line restored to fertility by the nuclear gene Fr2. However, steady-state levels of the 7.0- and 6.8-kb transcripts were decreased in the restored line relative to the CMS line.

DNA, Mitochondrial↗

Possible plasmid involvement in turimycin production in Streptomyces hygroscopicus.

Streptomyces hygroscopicus JA 6599 is the producer of the macrolide antibiotic turimycin. Mapping analysis by conventional matings and protoplast fusion techniques were carried out. The sequence of auxotrophic markers determined by using the method of minimizing the frequency of quadruple crossover recombinants, could be shown to be in accordance with the related marker sequence of Streptomyces coelicolor after both conjugation and protoplast fusion. However, the tur locus could localized between chromosomal markers only assuming quadruple crossover. Moreover, after conventional crosses the tur marker has to be localized at quite another site than after protoplast fusion. Regarding also our results on the evidence of extrachromosomal DNA in strains of S. hygroscopicus, the following hypothesis is proposed: the structural genes for turimycin biosynthesis are localized on the bacterial genome, but plasmid-borne genes might be involved in the control of the antibiotic production in a yet unknown way, possibly by inducing chromosomal rearrangements.

DNA, Bacterial↗

Chromosomal penicillin resistance in Staphylococcus aureus strains of phage group II.

The markers coding for serotype B penicillinase (PcB) in wild type strains of group II of S. aureus behaved as chromosomal characters in 6 out of 7 cases. The restriction deficient mutants also to some strains of group I. No extrachromosomal DNA could be detected in the transductants. The low frequency of transduction of chromosomal markers as well as the high restriction barrier seem to limit the spread of these markers outside group II.

Bacteriophage Typing↗

The maize plastid psbB-psbF-petB-petD gene cluster: spliced and unspliced petB and petD RNAs encode alternative products.

The chloroplast psbB, psbF, petB, and petD genes are cotranscribed and give rise to many overlapping RNAs. The mechanism and significance of this mode of expression are of interest, particularly because the accumulation of the psb and pet gene products respond differently to both light and, in C4 species such as maize, developmental signals. We present an analysis of the maize psbB, psbF, petB, and petD genes and intergenic regions. The genes are organized similarly in maize (a C4 species) and in several C3 species. Functional class II-like introns interrupt the 5' ends of petB and petD. Both spliced and unspliced RNAs accumulate; these encode alternative forms of the petB and petD proteins, differing at their N-termini. Promoter-like elements between psbF and petB, and biased codon usage suggest that the differential regulation of the psb and pet genes might be achieved at both the transcriptional and translational levels.

Amino Acid Sequence↗

Sequence and evolution of the regions between thr rrn operons in the chloroplast genome of Euglena gracilis bacillaris.

The rRNA genes are arranged in three sequential operons preceded by a fourth partial operon. Part or all of a 1462 nucleotide sequence extending from within the 3'-end of the 23S rRNA gene, across the 5S rRNA gene and a presumptive transcription terminator, to within the first structural gene (for 16S rRNA) of the rrn operon was determined for each region between operons. Homologies of the 3'-end of the 23S rRNA gene with the 4.5S rRNA genes of higher plant chloroplasts, and of the 5S rRNA gene with other 5S rRNA genes were examined. The region preceding the 16S rRNA gene, which is expected to contain sites for initiation and regulation of rrn transcription, includes a 305 base-pair sequence with substantial homology with structural genes elsewhere in the chloroplast genome. The homologies suggest that this portion of the leader evolved from copies of parts of the structural genes which had been inserted before the 16S rRNA genes. Thus the chloroplast rrn leader may provide a unique opportunity to study how a regulatory sequence evolved from well-defined structural genes.

Base Sequence↗

Role of the tof gene in the production and perpetuation of the lambdadv plasmid.

A series of lambda derivatives carrying tof mutations were tested for their ability to give rise to plasmid lambda dv. Phages carrying tof mutations that distorted expression of the pRoR-tof-OP operon, were unable to produce lambda dv. Phages carrying an altered tof gene, having only a moderate effect on the same operon, produced unstable lambdadv's. On the other hand, those tof mutants were only the expression of the pLoL-N-exo operon, but not that of the pRoR-tof-OP operon was affected, produced stable lambdadv's.

Coliphages↗

The generation of a ColE1-Apr cloning vehicle which allows detection of inserted DNA.

A 3.2 Mdal sequence of DNA, TnA, which contains the ampicillin (Ap) resistance determinant has been translocated from an R plasmid to the plasmid ColE1. A total of 12 isolates were studied. There are at least 8 sites in ColE1 at which TnA has inserted. Insertion at five of these has resulted in a Col-phenotype. One ColE1-Apr plasmid, RSF2124, was examined further and its replication properties are found to be similar to that of the parent plasmid. RSF2124 appears to be a useful plasmid vehicle for the molecular cloning of DNA from diverse prokaryotic sources: it codes for readily detectable Ap resistance and contains a single EcoRI site in a gene affecting colicin biosynthesis so that it is unable to produce colicin upon ligation to other DNA.

Ampicillin↗