Cell-free transcription of the nifH1 gene of Methanococcus thermolithotrophicus indicates that promoters of archaeal nif genes share basic features with the methanogen consensus promoter

Gohl, Harald Peter; Hausner, Winfried; Thomm, Michael

doi:10.1007/bf00279802

Cited by 13 publications

(3 citation statements)

References 31 publications

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“…Hence, this element is most likely specific for this promoter. Mutational studies of Methanococcus nifHl promoter that directs expression of a subunit of an archaeal nitrogenase [41] and of Pyrococcus GDH promoter [38], confirmed the importance of a TATA box for promoter function.…”

Section: Promoter Signalsmentioning

confidence: 83%

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Archaeal transcription factors and their role in transcription initiation

Thomm

1996

FEMS Microbiology Reviews

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Archaeal RNA polymerases show a weak ability in vitro to bind to promoter DNA and/or to initiate transcription with low activity independent of upstream regulatory DNA sequences. Active transcription in vitro and in vivo, however, depends strictly on a TATA box resembling the TATA box of eucaryal polII promoters. This TATA box is recognized by a polypeptide related to eucaryal TATA-binding protein (TBP) that was formerly designated aTFB. Template competition studies showed that this archaeal TATA-binding protein (aTBP) is stably sequestered at the promoter by interaction with the second archaeal transcription factor, aTFA, which is related to eucaryal transcription factor IIB (TFIIB). The association of archaeal TFIIB (aTFIIB) with the aTBP-promoter complex leads to template commitment, indicating that aTFIIB recruits archaeal RNA polymerase to the preinitiation complex. These analyses suggest the following order for assembly of transcription factors on the archaeal promoter: aTBP, aTFIIB, RNA polymerase, and provide evidence for a common molecular mechanism of transcription initiation by eucaryal RNA polymerase II and archaeal RNA polymerases. The sequence of the genes encoding aTBP and aTFIIB (TFB) showed all the characteristics conserved in their eucaryal counterparts. The degree of sequence similarity between archaeal and eucaryal transcription factors is between 27 to 35% for TFIIB and between 36 to 41% for TBP. The findings discussed here indicate that TBP and TFIIB perform analogous functions in Archaea and Eucarya and show that four essential components of archaeal and eucaryal transcriptional machineries. RNA polymerase, TATA box, TBP and TFIIB are homologous.

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Section: Promoter Signalsmentioning

confidence: 83%

“…cus [41] and of the GDH promoter of Pyrococcus furiosus [38] were analysed. Beside these in vitro studies, a tRNA Lys promoter of Haloferax volcanii was analysed in vivo using a transcriptional reporter system [42].…”

Section: Promoter Signalsmentioning

confidence: 99%

Archaeal transcription factors and their role in transcription initiation

Thomm

1996

FEMS Microbiology Reviews

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“…Comparison of the consensus sequence with results from other studies Previous studies applied site-directed mutagenesis to characterize promoters using in vitro transcription sytems from S. shibatae (Hain et a/., 1992; and M. vanielii (Hausner et al, 1991), and an in vivo sytem from H. volcanii (Palmer and Dennis, 1995). All but one of these studies (Gohl et a/., 1992) used stable RNA promoters. The resulting consensus sequences of three studies are included in Fig.…”

Section: Comparison Of the Consensus Sequence And Sequences Upstream mentioning

confidence: 99%

Characterization of the distal promoter element of halobacteria in vivo using saturation mutagenesis and selection

Danner

Soppa

1996

Molecular Microbiology

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The sequence and spacing requirements of the archaeal "distal promoter element' (DPE) were examined by randomizing positions -19 to -32 upstream of the transcriptional start site of the ferredoxin (fdx) promoter of Halobacterium salinarium. This randomized promoter library containing 4(14) entries was cloned in front of the dihydrofolate reductase (DHFR) reporter gene and transformed into Haloferax volcanii. Two approaches were used to characterize these synthetic promoters. First, 1040 independent clones were randomly chosen and their degrees of trimethoprim resistance were determined. The sequences of 20 clones that were either sensitive, partially resistant or very resistant, respectively, were determined. Secondly, the transformed library was screened by direct selection for high-activity promoters by growing transformants in the presence of trimethoprim. Both approaches produced the following consensus sequence for a halobacterial promoter: (Formula: see text) (where R = A or G; Y = C or T; W = A or T; S = G or C; N = A, C, G or T). Further characterization of two sensitive, two partially resistant, and two very resistant clones verified that DHFR activity and cell phenotype are directly correlated. Sensitive clones did not contain detectable dhfr mRNA, whereas partially resistant clones contained a 700 nucleotide (nt)-long transcript, and very resistant clones contained both the 700nt-long transcript and a second, more abundant, 500nt-long truncated transcript. Quantification of the dhfr mRNA and DHFR enzyme activity suggests that the 3'-untranslated region of the dhfr transcript, missing from the shorter transcript, functions as a negative regulator of translation.

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Structure and Organization of Genes

Reeve¹

1993

Methanogenesis

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Cell-free transcription of the nifH1 gene of Methanococcus thermolithotrophicus indicates that promoters of archaeal nif genes share basic features with the methanogen consensus promoter

Cited by 13 publications

References 31 publications

Archaeal transcription factors and their role in transcription initiation

Archaeal transcription factors and their role in transcription initiation

Characterization of the distal promoter element of halobacteria in vivo using saturation mutagenesis and selection

Structure and Organization of Genes

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