Upstream activating sequence

About: Upstream activating sequence is a research topic. Over the lifetime, 1633 publications have been published within this topic receiving 100112 citations.

Identification of enhancer elements in the upstream region of the nuclear photosynthetic gene ST-LS1.

Abstract: The nuclear gene ST-LS1 from potato encodes a 10-kilodalton protein that is a component of the oxygen-evolving complex of photosystem II. Analysis of the expression of a reporter gene driven by chimeric promoters, consisting of ST-LS1 upstream sequences and a truncated cauliflower mosaic virus 35S promoter, suggests that a strong positive regulatory element is located between position -345 and -261, whereas both the region -261 to +11 and the more upstream region -1600 to -530 are devoid of autonomous strong positive elements detectable by this approach. The ST-LS1 upstream region contains redundant elements conferring light-regulated and organ-specific expression, one of them being located between position -130 and +11. In addition, enhancer-like sequences conferring light-regulated as well as organ-specific expression to heterologous promoters were identified. These sequences are functional not only when located 5'-upstream of the coding region but also when placed 3'-downstream of the polyadenylation signal, thus representing one of the first examples of a plant gene-derived enhancer being able to induce a truncated heterologous promoter from a position 3'-downstream of the transcription unit.

Analysis of the upstream activating sequence and site of carbon and nitrogen source repression in the promoter of an early-induced sporulation gene of Bacillus subtilis.

Abstract: The transcription from the spoVG promoter of Bacillus subtilis is induced at the start of the stationary phase of growth and is dependent on the expression of the spoOA, spoOB, and spoOH genes. It is repressed in cells grown in the presence of excess glucose and glutamine and is under the negative control of the abrB gene. The spoOA and spoOB gene products function to suppress the negative control exerted by abrB. Transcription initiation requires the form of RNA polymerase holoenzyme that contains the spoOH gene product, sigma H. Optimal transcription also requires an upstream A-T-rich region termed the upstream activating sequence (UAS). The mechanism of UAS function was examined through mutational analysis of the spoVG promoter region. Deletion of the UAS or positioning the UAS one half turn or one full turn of the DNA helix upstream of its location in wild-type spoVG resulted in a severe reduction in promoter activity. Deletion of most of the UAS abolished the abrB-dependent repression of spoVG transcription. Higher activity was observed when the UAS was inserted 10 bp (one turn of the helix) upstream than when the sequence was repositioned either 5 or 13 bp upstream. Sequences upstream of the UAS were found not to be involved with the position-dependent function of the UAS. Positioning the UAS 42 or 116 bp upstream eliminated the stimulatory effect of the sequence on spoVG transcription. These data indicate that the UAS functions effectively when it is in close proximity to the -35 region. In vitro transcription analysis indicated that the deletion and insertion mutation affecting the UAS impair RNA polymerase-spoVG promoter interaction. Deletion of the UAS showed that the negative effect of exogenous glucose and glutamine is not dependent on the UAS but is exerted at a site within or near the -35 and -10 regions.

Sites of RNA polymerase III transcription initiation and Ty3 integration at the U6 gene are positioned by the TATA box.

Abstract: The function of a TATA element in RNA polymerase (EC 2.7.7.6) III transcription of a naturally TATA-containing U6 snRNA gene and a naturally TATA-less tRNA gene was probed by transcription and Ty3 transposition analyses. Deletion of the TATA box from a U6 minigene did not abolish transcription and Ty3 integration but changed the positions of initiation and insertion. Insertion of the U6 TATA box at three positions upstream of the TATA-less SUP2 tRNA(Tyr) gene resulted in novel transcription initiation and Ty3 integration patterns that depended upon position of the insertion. Nevertheless, the predominant tRNA gene initiation sites were not affected by insertion of the TATA sequence and remained at a fixed distance from the internal box A promoter element. Insertions of the TATA box upstream of a SUP2 box A mutant affected the level of transcription and restricted the use of upstream start sites, but they neither enhanced the use of TATA-dependent initiation sites nor restored expression to the level of the wild-type gene. We conclude that (i) the U6 TATA box is essential in vivo for correct initiation but not for transcription, (ii) a TATA box does not compensate for a weak box A sequence and so cannot perform equivalently, and (iii) the TATA-binding protein, and probably components of transcription factor IIIB, are present on the target at the time of Ty3 integration.

Identification of a novel TA-rich DNA binding protein that recognizes a TATA sequence within the brain creatine kinase promoter

Abstract: The rat brain creatine kinase gene possesses a structurally complex promoter with multiple potential regulatory elements. Two CCAAT sequences, a TATAAATA sequence and a TTAA sequence are found within the first one hundred base pairs. We present evidence that favors the allocation of the downstream TTAA sequence as the potential TATA box. We show that the CCAAT sequences and the upstream TATAAATA sequence are binding sites for potential regulatory factors and that sequences in this region are capable of regulating expression from the downstream TTAA sequence. We suggest that the protein that binds to the upstream TATAAATA sequence is not a classical TFIID factor but rather may serve to block the binding of TFIID and/or to promote transcription from the downstream start site. We have been able to define conditions in vitro under which binding to this upstream TATAAATA sequence does not occur. Under these conditions we are able to detect transcription from both potential TATA sequences, a situation which we have been unable to detect in vivo. Our experiments suggest the existence in HeLa and brain nuclei of a protein that recognizes the concensus TATAAATA sequence, that is distinct from TFIID, and that may function in part to deny access of TFIID to this potential promoter element.

A nucleosome-positioning sequence is required for GCN4 to activate transcription in the absence of a TATA element.

Abstract: In the gal-his3 hybrid promoter his3-GG1, the yeast upstream activator protein GCN4 stimulates transcription when bound at the position normally occupied by the TATA element. This TATA-independent activation by GCN4 requires two additional elements in the gal enhancer region that are distinct from those involved in normal galactose induction. Both additional elements appear to be functionally distinct from a classical TATA element because they cannot be replaced by the TFIID-binding sequence TATAAA. One of these elements, termed Q, is essential for GCN4-activated transcription and contains the sequence GTCAC CCG, which overlaps (but is distinct from) a GAL4 binding site. Surprisingly, relatively small increases in the distance between Q and the GCN4 binding site significantly reduce the level of transcription. The Q element specifically interacts with a yeast protein (Q-binding protein [QBP]) that may be equivalent to Y, a protein that binds at a sequence that forms a constraint to nucleosome positioning. Analysis of various deletion mutants indicates that the sequence requirements for binding by QBP in vitro are indistinguishable from those necessary for Q activity in vivo, strongly suggesting that QBP is required for the function of this TATA-independent promoter. These results support the view that transcriptional activation can occur by an alternative mechanism in which the TATA-binding factor TFIID either is not required or is not directly bound to DNA. In addition, they suggest a potential role of nucleosome positioning for the activity of a promoter.