Multiple protein - DNA interactions in the TATAA-less mouse thymidylate synthase promoter

Abstract

The mouse thymidylate synthase (TS) promoter lacks a TATAA box and directs transcriptional initiation at multiple sites over a 60 nucleotide region. All of the sequences that are important for transcription are located within close proximity to the first initiation site. Gel mobility shift and footprinting analyses with various sequences from this region of the TS promoter identified three major protein-DNA interactions. One of these corresponds to Sp1 interacting with a nonconsensus binding site downstream of the first transcriptional initiation site. Inactivation of the binding site by site-directed mutagenesis led to a 3-fold reduction in gene expression as well as a significant change in distribution of transcriptional start sites. The proteins responsible for the other two complexes (CII and CIII) do not appear to correspond to any of the common transcription factors that have been studied previously. The CII protein binds very close to the first initiation site. Inactivation of the binding site by site-directed mutagenesis had little effect on expression. The CIII protein binds immediately upstream of CII. Inactivation of this site led to a 12-fold reduction in expression, indicating that it is important for expression of the TS gene.

Regulation of the human poly(ADP-ribose) polymerase promoter by the ETS transcription factor

“…Moreover, ETS motifs are frequently located in the vicinity of the transcription start site in a number of promoters that lack a TATA element, including the genes encoding DNA polymerase a and b (Pearson et al, 1991;Yamaguchi et al, 1988), thymidilate synthetase (Jolli and Johnson, 1991), von Willebrand factor (Schwachtgen et al, 1998), Ets2 (Mavrothalassitis et al, 1990, the tumor suppressor gene Maspin (Zhang et al, 1997) and number of myeloid speci®c genes (reviewed by Moreau-Gachelin, 1994). It seems that PARP can be added to this group of genes because it exhibits features typical of TATA-de®cient promoters and has consensus ETS motifs close to the major initiation sites.…”

Promoter alteration causes transcriptional repression of the POMGNT1 gene in limb-girdle muscular dystrophy type 2O

“…19 In many TATA-less promoters, Sp1 binding is required to determine the transcription start site. 20 Another transcription factor potentially implicated in the transcriptional regulation of POMGNT1 gene is Ets1, which binds to the core (5¢-GGAA/T-3¢) sequence. Our EMSA assay detected one Ets1-binding site located in the +2 to +8 bp region.…”

GA-binding protein factors, in concert with the coactivator CREB binding protein/p300, control the induction of the interleukin 16 promoter in T lymphocytes

“…Many TATA-less RNA polymerase II promoters contain typical initiator elements, i.e., short conserved sequences that are bound by proteins of the transcriptional machinery. Some of these promoters, including the CD3, CD4, CD11c, CD18, CD72, IL-2R␤, TGF␤ RII, Ets-2, murine laminin B2, type IV collagenase, thymidylate synthetase, and cytochrome c oxidase subunit IV promoters possess Ets-like binding motifs near their transcriptional start sites, suggesting a new class of initiator elements (34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45). Recently, two PEA3 motifs that are bound by GABP factors were shown to function as a minimal transcriptional initiator element when placed in front of a luciferase gene (46).…”