Sequence-specific interactions of a pea nuclear factor with light-responsive elements upstream of the rbcS-3A gene.

Abstract

Pea nuclear extracts were used in gel retardation assays and DNase I footprinting experiments to identify a protein factor that specifically interacts with regulatory DNA sequences upstream of the pea rbcS‐3A‐gene. This factor, designated GT‐1, binds to two short sequences (boxes II and III) in the −150 region that are known to function as light‐responsive elements (LREs) in transgenic tobacco. Binding of GT‐1 to homologous sequences further upstream (boxes II and III in the −220 region) indicates that these boxes comprise the redundant LRE that functions in vivo when boxes II and III are deleted. In both box II and box II, methylation interference experiments demonstrate that two adjacent G residues are critical for GT‐binding. Single Gs present in boxes III and III are also important. Since GT‐1 is present in nuclear extracts from leaves of light‐grown and dark‐adapted pea plants, its regulatory role does not depend on de novo synthesis. Thus if GT‐1 binds differentially in vivo it must be postranslationally modified or sterically blocked from binding by another factor in response to light.

PLANT HOMOLOGOUS TO PARAFIBROMIN Is a Component of the PAF1 Complex and Assists in Regulating Expression of Genes within H3K27ME3-Enriched Chromatin

“…Arabidopsis nuclear proteins were prepared using a modification of the methods described previously (Green et al, 1987;Bowler et al, 2004). Briefly, 50 g of inflorescences including stems and young siliques were collected, rinsed with prechilled water, and homogenized in 200 mL of ice-cold buffer A (1 M hexylene glycol, 10 mM PIPES/KOH [pH 7.0], 10 mM MgCl 2 , and 0.2% Triton X-100, containing 5 mM 2-mercaptoethanol, 1 mM phenylmethylsulfonyl fluoride, and 0.1 mg/mL pepstatin A).…”

Isolation and characterization of strong gene regulatory sequences from apple, Malus × domestica

“…2), which are known for their light responsive activity (Castresana et al 1988). Also an element showing similarity to a GT-1 binding box II, which was identified in the promoter of pea RbcS-3A (Green et al 1987), is present in the MdRbcS promoter. This element is associated with high expression levels of RbcS-3A in pea.…”

Interaction of nuclear factors with upstream sequences of a lipid body membrane protein gene from carrot.

“…This has also been found in other plant systems (Maier et al, 1987;Deikman and Fisher, 1988). Recently, it has been shown that multiple cis-elements result in several complexes with nuclear factors in animal (Babiss et al, 1987;Celeghini et al, 1987;Lichtsteiner et at., 1987;Monaci et al, 1988) as well as in plant systems (Green et al, 1987;Deikman and Fischer, 1988), and it has been proposed that enhancer-like elements modulate transcription although they are located relatively far upstream of their respective genes. It is interesting to note that the region of fragment 111 responsible for the formation of the C1 complex contains the sequence TGTGGATATG, which is similar to a motif found nOt only in many vira1 enhancers (Odell et al, 1985;Sassone-Corsi and Borrelli, 1986) but also upstream of several plant genes (Timko et al, 1985;Kaulen et al, 1986;Maier et al, 1987).…”

“…Embryo and callus nuclear extracts were isolated from somatic embryo and callus cultures according to Green et al (1987). Endlabeled and competitor DNA fragments were isolated by phenol extractions atter electrophoresis in low-melting-point agarose.…”