Transcription initiation of the Saccharomyces cerevisiae iso-1-cytochrome c gene

McNeil, J B; Smith, Margaret Caroline MacHin

doi:10.1016/0022-2836(86)90439-0

Cited by 120 publications

(79 citation statements)

References 36 publications

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“…In higher eucaryotes, the distance between the TATA box and the initiation site is fixed at about 30 bp (2). This is different from the situation in yeasts in which the distance between the TATA box and the initiation site is both larger and variable (6,17,31,35,42). Our studies agree with these findings; the CYC7 TATA sequence directed transcription to start at sites greater than 45 bp downstream, with a great deal of flexibility as to the exact distance.…”

Section: Discussionsupporting

confidence: 67%

Sequences required for transcriptional initiation of the Saccharomyces cerevisiae CYC7 genes.

1987

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A series of BAL 31 deletions were constructed in the upstream region of the Saccharomyces cerevisiae CYC7 gene to determine sequences required for transcriptional initiation. These deletions identified the TATA box as an alternating A-T sequence at -160 and the initiation sequences as well as the spatial relationship between them. The TATA box was necessary for wild-type levels of expression of the CYC7 gene. Decreasing the distance between the TATA sequence and the initiation site did not alter gene expression, but the site of transcription was shifted 3'-ward. In most cases, transcription initiated at a number of sites, the 5'-most of which was the first suitable site greater than 45 base pairs 3' of the TATA sequence, suggesting a spatial relationship between these sequences, Consensus sequences previously proposed for initiation sites were evaluated with respect to the start sites identified in this study as well as the start sites of other yeast genes.The selection of the initiation site for the transcription of class II genes of higher eucaryotes is governed by an A+T-rich sequence called the TATA box which directs RNA polymerase II to begin transcription 30 base pairs (bp) downstream (2). Deletion of a TATA sequence results in either decreased gene expression, initiation at novel sites, or both (1,9,28,30). If the distance between the TATA sequence and the initiation site is altered, transcription begins at a new location 30 bp downstream from the relocated TATA sequence (1). No sequence preference at the start site is evident, although transcription often begins at an A residue.In Saccharomyces cerevisiae, TATA-like sequences have been identified in the upstream region of most class II genes and are thought to play a role in transcriptional initiation (45). However, for several reasons it is difficult to assess their importance by inspection of the sequence alone. For one, the regions upstream of yeast genes are generally A+T-rich, containing many candidates for TATA boxes. Second, many yeast genes have multiple start sites, making the assignments of TATA boxes to start sites difficult. Finally, in those cases in which the TATA sequence has been studied, there is not a clear relationship between the spacing between the TATA box and the initiation site.Recent studies of the CYC1 (17), HIS3-DEDI (6), HIS4 (35), and PH05 (42) genes have defined a range over which TATA sequences can act; deletions altering the spacing between the TATA sequence and the initiation site point to an effective range of 40 to 120 bp over which TATA sequences can direct transcription. Given the variability in the distance between TATA sequences and the initiation site, it follows that signals contained in sequences at the initiation site must play a role in determining where transcription should begin (4,6,13,17,32,35,42). However, the different studies have identified different sequences. Clearly, more data are required to carry out any comprehensive comparative study.The S. cerevisiae CYC7 gene codes for 5% of the cytochrome c pr...

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Section: Discussionsupporting

confidence: 67%

Sequences required for transcriptional initiation of the Saccharomyces cerevisiae CYC7 genes.

1987

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“…After the addition of ethanol, the mixture was kept at -20°C overnight, and the sediment was washed with ethanol (70%, by vol.). The dried sediment was treated as described by McNeil and Smith [38]. However, the reaction mixture contained 1 mM each of dGTP, dCTP and dTTP, and 1 pM 35S-labelled deoxyadenosine-5'-[a-thio] triphosphate (1.2 Ciipmol).…”

Section: Methodsmentioning

confidence: 99%

Structure and Function of a Second Gene Cluster Encoding the Formate Dehydrogenase of Wolinella Succinogenes

Lenger

Herrmann

Groß

et al. 1997

European Journal of Biochemistry

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Wolinella succinogenes contains a single formate dehydrogenase, but two gene loci (fdhl and fdhII) code for the subunits of the enzyme. The nucleotide sequence of fdhII is almost identical with that of fdhI in the region comprising fdhEABCD. The sequences of fdhI and fdhll differ in the promotor regions upstream of fdhE. Deletion mutants lacking either fdhI or fdhII synthesize functional formate dehydrogenases, as shown by growth with formate as electron donor and either fumarate or polysulfide as acceptor substrates, and by the presence of the FdhA subunit and of enzyme activity. In the wild-type strain, the fdhl genes appear to be expressed preferentially during growth with formate and fumarate. The six-times greater amount of the enzyme present upon growth with formate and polysulfide is due to the expression of both fdhI and fdhll. In a previous publication [4], the fdhA gene (of the fdhI cluster) was cloned from a genomic library of W. succinogenes using an antiserum raised against FdhA. The fdhA gene preceded the genes fdhB, fdhC and a fourth open reading frame (fdhD), the function of which is not known. The gene fdhA is predicted to encode a prepeptide of the catalytic subunit with an N-terminal signal peptide (32 residues) typical of certain periplasmic proteins or membraneous proteins facing the periplasm [15]. Consistently, the substrate site of the enzyme is oriented towards the periplasmic side of the membrane [6]. While the genes encoding the other electron-transport enzymes (fumarate reductase [16], hydrogenase [ 171 and polysulfide reductase [9]) were found only once in the genome of W succinogenes, there was evidence indicating that a functional formate dehydrogenase could be formed in the absence of the fdhl genes (41. Therefore, the second gene locus (fdhll) was sequenced, and the functions of fdhI and fdhlI were investigated using a AfdhI and a AfdhlI mutant. MATERIALS AND METHODSPhage, bacterial strains, plasmids and growth conditions. Propagation of phage AEMBL3 [18]

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“…To create pSK113, pGAL4CG-(23) was digested with Xho I and Sma I, and the liberated CYCI promoter-G-free cassette fusion was cloned into the Xho I and Xba I (blunt-ended by the Klenow fragment) sites of pGEM-7Zf(+) (Stratagene). The 5' end of the CYCI promoter of pSK113 is nucleotide position -138 with respect to the translation initiation site at +1 (24). pSK114 was constructed by replacing the Xho I-Ava II region (from -138 to -68) of pSK113 with the Xho I-Ava II fragment (from -248 to -68) ofthe authentic CYCI promoter (24).…”

Section: Methodsmentioning

confidence: 99%

“…The 5' end of the CYCI promoter of pSK113 is nucleotide position -138 with respect to the translation initiation site at +1 (24). pSK114 was constructed by replacing the Xho I-Ava II region (from -138 to -68) of pSK113 with the Xho I-Ava II fragment (from -248 to -68) ofthe authentic CYCI promoter (24). Two (25) has essentially the same construction to pSK115, except that pJJ460 carries a shorter G-free sequence.…”

Section: Methodsmentioning

confidence: 99%

Yeast GAL11 protein is a distinctive type transcription factor that enhances basal transcription in vitro.

Sakurai

Hiraoka

Fukasawa

1993

Proc. Natl. Acad. Sci. U.S.A.

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The yeast auxiliary transcription factor GAL1l, a candidate for the coactivator, was partially purified from yeast cells, and its function was characterized in a cell-free ranscription system. The partially purified GAL1l protein stimulated basal transcription from the CYCI core promoter by a factor of 4-5 at the step of preinitiation complex formation. GALll protein also enhanced transcription activated by general regulatory factor 1, GAL4-AH, or GAL4-VP16 to the same extent as the basal ranscription. Therefore, the apparent potentiation of the activators by GALll was attributable to the stimulation of basal transcription. The wild-type GALll protein (but not a mutant-type protein) produced in bacteria stimulated transcription as effectively as GAL1l from yeast. These results suggest that GALll functions as a positive cofactor of basal and activator-induced transcription in a cell-free transcription system. that potentiates a weak activator GAL4-AH. Surprisingly, the mutation is a missense mutation within GALI and was therefore designated GALIIP (P stands for potentiator). They suggest a direct interaction between GAL1l and GAL4 molecules and suggest that the complex of the two proteins is a strong activator. They further find that GALII is required for normal functioning of PPR1, an activator for the URA3 gene. Puzzlingly, GALI is also involved in the transcriptional repression of genes, since a loss-of-function mutation of GALII, called sptl3, restores the transcription of genes inactivated by insertion of the yeast transposon Ty (22).To get insight into the complex pleiotrophic functions of GALH in vivo, we analyzed the role of GAL1l biochemically. We have partially purified GALli protein and studied how it works in a cell-free transcription system. We have found that GAL1l enhances the basal transcription as well as the transcription activated by GAL4-AH, GAL4-VP16, or GRF1. These results suggest that, since GAL1l regulates the basal transcription, it is not a bona fide coactivator, but rather it belongs to a distinctive class of basal transcription factors. (15,27). The major start site of GALli mRNA was Abbreviations: UAS, upstream activation sequence; GRF1, general regulatory factor 1; GST, glutathione S-transferase. MATERIALS AND METHODSITo whom reprint requests should be addressed. 8382

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Transcription initiation of the Saccharomyces cerevisiae iso-1-cytochrome c gene

Cited by 120 publications

References 36 publications

Sequences required for transcriptional initiation of the Saccharomyces cerevisiae CYC7 genes.

Sequences required for transcriptional initiation of the Saccharomyces cerevisiae CYC7 genes.

Structure and Function of a Second Gene Cluster Encoding the Formate Dehydrogenase of Wolinella Succinogenes

Yeast GAL11 protein is a distinctive type transcription factor that enhances basal transcription in vitro.

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