A protein factor, identified in nuclear extracts obtained from tomato (Lycopersicon esculentum, Solanaceae) and Arabidopsis thaliana (Brassicaceae) seedlings, specifically binds upstream sequences from the plant light-regulated gene family encoding the small subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase (RBCS). RBCS upstream sequences from tomato, pea (Pisum sativum, Leguminosae), and Arabidopsis are recognized by the factor. The factor recognition occurs via a short conserved sequence (G box) whose consensus sequence is 5'-TCTTACACGTGGCAYY-3' (where Y is pyrimidine). This sequence is distinct from the GT motif described previously in RBCS promoters. Two other conserved sequences, showing a lesser degree of evolutionary conservation, are found upstream of the G box but do not bind to the G box binding factor (GBF). Twelve nucleotides within the G box are sufficient for the formation of a stable DNA-GBF complex. GBF is found in both light-grown and dark-adapted tomato leaf extracts, but it is present in greatly reduced amounts in root extracts.

https://www.pnas.org/content/pnas/85/19/7089.full.pdf

An evolutionarily conserved protein binding sequence upstream of a plant light-regulated gene.

The nucleotide sequence of the lac promoter-operator region has been determined. The 122 base pairs comprising this region include the recognition sites for RNA polymerase, the positive regulatory protein, CAP, and the negative regulatory protein, the repressor. Identification of mutant variants of the sequence combined with the in vitro biochemical studies of others has allowed us to tentatively identify the recognition site for each of these proteins, and to suggest how CAP might act at a distance to affect the interaction of RNA polymerase with the promoter.

https://science.sciencemag.org/content/sci/187/4171/27.full.pdf

Genetic regulation: the Lac control region.

This review will attempt to cover the present information on the multiple forms of eukaryotic DNA-dependent RNA polymerases, both at the structural and functional level. Nuclear RNA polymerases constitute a group of three large multimeric enzymes, each with a different and complex subunit structure and distinct specificity. The review will include a detailed description of their molecular structure. The current approaches to elucidate subunit function via chemical modification, phosphorylation, enzyme reconstitution, immunological studies, and mutant analysis will be described. In vitro reconstituted systems are available for the accurate transcription of cloned genes coding for rRNA, tRNA, 5 SRNA, and mRNA. These systems will be described with special attention to the cellular factors required for specific transcription. A section on future prospects will address questions concerning the significance of the complex subunit structure of the nuclear enzymes; the organization and regulation of the gene coding for RNA polymerase subunits; the obtention of mutants affected at the level of factors, or RNA polymerases; the mechanism of template recognition by factors and RNA polymerase.

Eukaryotic RNA polymerases.

The experimental description of the mechanics as well as the regulation of bacterial protein synthesis appears to have entered a final stage. Virtually every step involved in the assembly of amino acids into protein-chain initiation, elongation, termination-can be performed in the test tube wi-th components purified from bacterial cells (1). The steps in bacterial messenger RNA (mRNA) biogenesis are almost as well understood. The functional RNA polymerase and several regulatory proteins have been isolated and shown to participate in the control of synthesis of specific mRNA, both under conditions where only RNA is produced and in coupled systems where all the steps occur between the initiation of mRNA production and completion of functional enzyme protein (1). These awesome biochemical achievements with bacterial preparations probably would not have been possible, certainly not at this time, if a decade of successful bacterial genetics had not provided evidence concerning the mode of regulation that exists-namely, control of transcription of mRNA by regulatory proteins. Presented with the challenge of understanding protein synithesis in mammalian cells in detail comparable to the I-ent understanding in bacteria, the ':eh biologist might, after a first appraisal, despair. The genetic methods for isolating and characterizing mutants which might affect the availability of mRNA do not exist for mammalian cells and, if developed, will certainly present greater practical difficulties in the-characterization and experimental

https://science.sciencemag.org/content/sci/181/4106/1215.full.pdf

Biogenesis of mRNA: Genetic Regulation in Mammalian Cells

https://www.cell.com/cell/pdf/0092-8674(92)90174-B.pdf

Polypeptides containing highly conserved regions of transcription initiation factor σ70 exhibit specificity of binding to promoter DNA

On the Rule of Different Subunits of DNA-Dependent RNA Polymerase from E. coli in the Transcription Process

The decrease in the activity of DNA-dependent RNA polymerase in Escherichia coli after infection with bacteriophage T4 is the consequence of a structural change of the polymerase and not of the appearance of an inhibitor. The kinetics of this change were followed using polymerases highly purified at different times after infection (a) by measuring the specific enzymatic activity with T4- and calf thymus DNA as templates and (b) by following the appearance of the modified polymerase-subunit in disc-electrophoresis.



The results allow one to differentiate between two consecutive processes:



1
A very rapid alteration which inactivates the RNA polymerase for T4-DNA, but not for other DNA templates, and which does not require protein synthesis.

2
A structural modification which depends on protein synthesis and leads to a change in one subunit of the polymerase.





As shown by the use of the T4-mutants am BL 292 and am N.134 neither gene 55 nor gene 33, both involved in the transcription of late genes, are participating in the modification reaction.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1969.tb00611.x

Kinetics of the Alteration and Modification of DNA‐Dependent RNA‐Polymerase in T4‐Infected E. coli Cells

DNA-dependent RNA polymerase from bacteriophage T4-infected Escherichia coli cells differs in all its subunits from the host's core enzyme.



T4-modified α differs from E. coliα by 1 or 2 additional peptides. One additional tyrosine positive peptide and the displacement of one histidine containing peptide is characteristic for T4-modified β'. T4-modified β is significantly different from E. coliβ in its histidine and tryptophan containing peptides, whereas differences in the tyrosine peptide pattern are less obvious.



No common features were observed among the peptide patterns of the subunits from E. coli RNA polymerase.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1971.tb01571.x

Fingerprint Maps of Tryptic Peptides from Subunits of Escherichia coli and T4‐Modified DNA‐Dependent RNA Polymerases

1
After infection with bacteriophage T4 the DNA-dependent RNA polymerase of the Escherichia coli host is changed in two consecutive steps: alteration and modification.

The alteration takes place immediately after phage adsorption and is independent of protein synthesis. It leads to the shut off of E. coli specific RNA synthesis and permits the transcription of the immediate early phage genes.

Under conditions allowing phage specific protein synthesis the E. coli core polymerase is modified. The first step of modification is completed within 4 min after phage infection (25°C). The purified modified enzyme does not contain σ-factor and exhibits structural changes in all the subunits of the core.

2
Under conditions which do not allow DNA synthesis (infection with T4 am N82) the net rate in vivo of immediate early, followed by delayed early gene transcription shows one early burst.

In normal phage development immediate early and delayed early transcription display a second rise preceding replication. Late RNA is made after the onset of DNA synthesis.

3
In crude extracts prepared from E. coli cells infected under conditions which allow as well as under conditions which do not allow protein synthesis, one burst of transcriptional capacity follows infection. The length of this burst is sufficient for the transcription of approximately one reading unit. Although transcription in vivo continues under conditions allowing protein synthesis, polymerase activity in crude extracts in vitro is very low after the burst even when exogenous template DNA is added.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1971.tb01572.x

A Correlation of Changes in Host and T4 Bacteriophage Specific RNA Synthesis with Changes of DNA‐Dependent RNA Polymerase in Escherichia coli Infected with Bacteriophage T4

Melitta Schachner

Papers

On the Rule of Different Subunits of DNA-Dependent RNA Polymerase from E. coli in the Transcription Process

Kinetics of the Alteration and Modification of DNA‐Dependent RNA‐Polymerase in T4‐Infected E. coli Cells

Fingerprint Maps of Tryptic Peptides from Subunits of Escherichia coli and T4‐Modified DNA‐Dependent RNA Polymerases

A Correlation of Changes in Host and T4 Bacteriophage Specific RNA Synthesis with Changes of DNA‐Dependent RNA Polymerase in Escherichia coli Infected with Bacteriophage T4