A. V. Soldatov

A review of the internet site practical molecular biology

In 2000, in the middle of September, the Russian Web site Practical Molecular Biology (http://molbiol.edu.ru) was opened. The main tasks of the project are (i) distribution of experimental work experience among Russian and foreign laboratories and (ii) organization of an information database in Russian for researchers connected with molecular biology or medicine.

The Involvement of the Drosophila melanogaster Nuclear Protein e(y)2 in Transcription Regulation

Molecular analysis of a new evolutionarily conserved transcription factor, e(y)2, was carried out. The protein was detected in a complex of approximately 700 kDa contained in aDrosophila melanogaster transcription nuclear extract. The e(y)2 protein was shown to interact with components of the preinitiation transcription complex TFIID. Addition of e(y)2 to a transcription extract of HeLa cells increased transcription 4–5 times when chromatin, but not free DNA, was used as a template. Genetic analysis showed that the C-terminal amino acid residues of transcription factor TAFII40 are important for its interaction with e(y)2.

Molecular Characterization of a New Evolutionarily Conserved Nuclear Protein e(y)2

Molecular structural and function analyses of the Drosophila melanogasterenhancer of yellow 2 (e(y)2) gene showed that its product acts as a transcription factor and is one of the basic elements of the eukaryotic transcription system. The gene is expressed at all stages of D. melanogaster development and consists of a single intron coding for the protein of 101 amino acid residues. The e(y)2 protein does not contain regions homologous to known proteins. The protein binds with chromatin but not with DNA. On evidence of immune staining, e(y)2 occurs in the nuclei of all D. melanogaster cells. Each nucleus contains approximately 1.2 × 104 molecules of the protein. Immune staining revealed approximately 200 sites of e(y)2 location on polytene chromosomes. The protein is evolutionarily conserved: its homologs were found in evolutionary distant organisms, such as plants, mammals, and protozoans. Amino acid sequences of human, rabbit, and mouse e(y)2 are identical to each other.

Molecular characterization of two newDrosophila melanogaster homologs of human TAFII30

Two new homologs of human (h) TAFII30, dTAFII16 and dTAFII24 were revealed inDrosophila melanogaster. The proteins are encoded by neighboring genes and bind with the TATA-binding protein and other dTAFII proteins involved in TFIID. Only dTAFII24 interacts with GCN5 histone acetyltransferase (HAT), which is the first demonstration of the TAFII-GCN5-HAT complex inD. melanogaster. The two proteins have both common and individual location sites on polytene chromosomes. Possibly, the functions of dTAFII16 and dTAFII24 are similar but not identical.

Millimolar concentrations of methylmethanesulfonate enhance transcription from the cytomegalovirus early promoter at least 100-fold

Treatment of cultured cells with methylmethanesulfonate enhanced transcription from the CMV early promoter 25–250 times and protein synthesis 60–100 times in a dose-dependent way. The effect was promoter-specific and was not observed with a retroviral LTR promoter. Structural analogs such as ethylmethanesulfonate, trifluoromethanesulfonate, andp-toluenesulfonate did not enhances expression from the CMV promoter.

In vivo analysis of theDrosophila melanogaster e(y)1/TAF II40 gene

A study was made of the molecular structure and properties of theDrosophila melanogaster enhancer of yellow 1 (e(y)1) gene which is involved in long-range promoter-enhancer interactions. The gene codes for transcription factor TAFII40 (a component of the pre-initiation transcription complex TFIID) and is expressed in all cells at all developmental stages. The protein occurs in virtually all discs of polytene chromosomes. Thee(y)1 gene is maternal, and its product is accumulated in oocytes. Suppression ofe(y)1 is lethal. Two mutations obtained,e(y)1u1 ande(y)1P1, result from insertion of Stalker and theP element, respectively. Genetic analysis ofe(y)1u1 suggests that internal promoters are sensitive to a decrease in TAFII40.