Tau factor from Escherichia coli mediates accurate and efficient termination of transcription at the bacteriophage T3 early termination site in vitro

Briat, Jean François; Bollag, Gideon; Kearney, Clare A.; Molineux, Ian J.; Chamberlin, Michael J.

doi:10.1016/0022-2836(87)90456-6

Cited by 18 publications

(14 citation statements)

References 30 publications

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“…This low efficiency of intrinsic termination by the purified enzyme suggests that accessory components of the transcription machinery modulate terminator utilization in the cell. It should be noted that there are a number of bacterial rho-independent termination sites that are very efficient in vivo yet are inefficient in vitro with the purified bacterial RNA polymerase in the absence of accessory factors (6,9,47).…”

Section: Resultsmentioning

confidence: 99%

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Kerppola¹,

Kane²

1988

Mol. Cell. Biol.

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We have studied transcription elongation and termination in the human c-myc gene. Transcription of c-myc gene sequences with purified mammalian RNA polymerase II revealed several sites of transcription termination and pausing in the vicinity of the exon 1-intron 1 junction. This region previously has been shown to block transcription elongation in vivo by nuclear run-on analysis (D. Bentley and M. Groudine, Nature [London] 321:702-706, 1986). These sites were recognized by purified RNA polymerase II, and we therefore designated them intrinsic sites of termination and pausing. Two of these sites cause termination of RNA polymerase Ill transcription as well. RNA polymerase II terminated transcription in a cluster of seven consecutive T residues in the nontranscribed strand and paused during transcription at three additional sites in this region. The intrinsic sites of transcription termination and pausing described here correspond closely to the 3' ends of transcripts synthesized in Xenopus oocytes injected with plasmids containing the c-myc termination region (D. Bentley and M. Groudine, Cell 53:245-256, 1988). This correspondence suggests that the intrinsic recognition of these termination and pause sites by purified RNA polymerase II may play a role in the transcription elongation block observed in vivo.

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Section: Resultsmentioning

confidence: 99%

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Kerppola¹,

Kane²

1988

Mol. Cell. Biol.

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“…The template for transcription was generated by PCR from pHA10O-R63 using the T3 and T7 universal primers. Template 2 was a 302 bp PCR fragment amplified from pAR1707 (Briat et of., 1987) which contains the bacteriophage T7 A I promoter. Primer end points relative to the A I transcription initiation site were -149 and + 153.…”

Section: Dna Templatesmentioning

confidence: 99%

Role of Escherichia coli RNA polymerase alpha subunit in modulation of pausing, termination and anti-termination by the transcription elongation factor NusA.

et al. 1996

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The alpha subunit (alpha) of RNA polymerase (RNAP) is critical for assembly of polymerase and positive control of transcription initiation in Escherichia coli. Here, we report that alpha also plays a role in transcription elongation, and this involves a direct interaction between alpha and NusA factor. During in vitro transcription without NusA, alpha interacts with the nascent RNA, as revealed by photocrosslinking. When NusA is present, RNA crosslinks to NusA rather than to alpha. We show that this NusA‐RNA interaction is diminished during transcription with an RNAP mutant that lacks the C‐terminus of alpha beyond amino acid 235, including the so‐called alpha CTD. The absence of alpha CTD also affects NusA's ability to enhance transcription pausing, termination at intrinsic terminators and anti‐termination by the phage lambda Q anti‐terminator, but not anti‐termination by the lambda N anti‐terminator. NusA functions are not recovered even when transcription with mutant RNAP is done with excess NusA, a condition which does restore NusA‐RNA crosslinking. By affinity chromatography, we show that NusA interacts directly with alpha, and also with beta and beta’, but not with mutant alpha. Hence, alpha‐NusA interaction is vital for the control of transcript elongation and termination.

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“…The early terminators of the bacteriophages T7 and T3 have very similar sequences (Briat et al, 1987; Figure 1). Both can be classified as p-independent terminators since they have typical p-independent terminator stem-loop and tail structures, 0006-2960/89/0428-5210S01.50/0 © 1989 American Chemical Society and neither is affected by p mutations (Kiefer et al, 1977).…”

mentioning

confidence: 97%

Terminator-distal sequences determine the in vitro efficiency of the early terminators of bacteriophages T3 and T7

Telesnitsky¹,

Chamberlin²

1989

Biochemistry

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Bacteriophages T3 and T7 contain homologous terminators for Escherichia coli RNA polymerase that restrict early phage transcription to the leftmost 20% of the linear phage genomes. These two terminators serve equally well as p-independent terminators in vivo, but their in vitro efficiencies and sensitivity to salt and nucleotide concentrations differ dramatically. Sequence analysis shows that the T7 and T3 terminators differ at only two sites in the region normally accepted as defining terminator function. In order to determine which structural features of these two terminators are responsible for their functional differences, a series of hybrid terminators were constructed in which structural features of the two terminators were systematically interchanged. Transcription of hybrid terminator templates revealed that sequences downstream of the termination release sites are responsible for the differences in efficiency of in vitro termination. These sequences also determine the sensitivity of these terminators to elevated salt concentrations and to alterations of substrate concentrations. Alteration of the sequences in the region between three and seven nucleotides downstream of the final T7Te release site is sufficient to reduce termination efficiency to that of T3Te, and point mutations in this region yield terminators with intermediate efficiency. Hence, the determinants of p-independent terminator efficiency in vitro must include elements of the transcription complex other than the structure of the 3' end of the transcript. The termination differences between T7Te, T3Te, and their hybrid derivatives are overcome in vivo; all of these sites become very efficient. This finding further supports the hypothesis that protein factors or other cellular features enhance the efficiency and specificity of p-independent terminators in vivo.

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Tau factor from Escherichia coli mediates accurate and efficient termination of transcription at the bacteriophage T3 early termination site in vitro

Cited by 18 publications

References 30 publications

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Role of Escherichia coli RNA polymerase alpha subunit in modulation of pausing, termination and anti-termination by the transcription elongation factor NusA.

Terminator-distal sequences determine the in vitro efficiency of the early terminators of bacteriophages T3 and T7

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