细胞技术

Steroid hormone receptors are members of the nuclear receptor family of ligand-activated transcriptional regulatory proteins. Recent work in the field of nuclear receptor action has demonstrated an association of receptors with coregulatory proteins termed “coactivator ...

The mouse mammary tumor virus (MMTV) promoter has been used extensively as a model system to examine the role of chromatin structure on transcriptional regulation from a steroid responsive gene (Fig. 1). Early studies demonstrated that the chromatin structure of the MMTV promoter was alter ...

The glucocorticoid, mineralocorticoid, progesterone, androgen, estrogen α and estrogen β receptors (GR, MR, PR, AR, ERα, and ERβ, respectively) form the steroid receptor family, part of the nuclear receptor superfamily (1). Like other nuclear receptors, steroid receptors have a conser ...

Biological effects of steroids are mediated through an initial interaction with specific receptor belonging to a member of the steroid/thyroid/retinoid receptor gene superfamily (1,2). Recently, increasing numbers of new members of this family, including their subtypes have be ...

Steroid hormones bind to their specific nuclear receptor protein, which are bound to their DNA receptor motifs and become activated to turn on transcription of other genes (1). As an important example, estrogen binds to estrogen receptors (ERs) and activates the progesterone receptor gene ...

Combinatorial chemistry has significantly increased the speed of small molecule lead discovery and optimization for multiple targets in the pharmaceutical industry. Large libraries of bioactive compounds, synthesized either as mixtures or discreet units, can now be made using ...

The MCF-7 cell line was isolated from a pleural metastasis of a human breast adenocarcinoma, and, when grown on plastic substrates, typically forms a continuous cell monolayer at confluence (1). MCF-7 cell cultures respond to 17β-estradiol (E2) by increases in the expression of a number of genes (( ...

The yeast two-hybrid system is a powerful tool for the isolation and characterization of nuclear receptor interacting proteins such as coactivator and corepressor proteins. Generally coactivators associate with the nuclear receptors in an agonist-dependent manner and this sp ...

The cell cycles of the budding yeast Saccharomyces cerevisiae and the fission yeast, Schizosaccharomyces pombe are currently the best understood of all eukaryotes. Studies in these two evolutionarily divergent organisms have identified common control mechanisms, which have p ...

This chapter reviews the basic features of plant cell proliferation. Although plant cell division seems to be largely similar to animals and fungi, there are a number of peculiarities that are perhaps related to their lifestyle and development. Initial comparisons of animal, fungal, and pl ...

Drosophila melanogaster provides an outstanding experimental system to study the regulation of cell cycle progression during animal development. Sophisticated forward and reverse genetic techniques and the ability to observe detailed cell biological phenomena in vivo ha ...

The nematode C. elegans provides an animal model that allows for genetic dissection of cell cycle regulation in the context of development. Processes such as progression through meiotic prophase, spindle function, chromosome segregation, and cytokinesis are attractive topics to s ...

Oocytes, eggs, and embryos from the frog Xenopus laevis have been an important model system for studying cell cycle regulation for several decades. First, progression through meiosis in the oocyte has been extensively investigated. Oocyte maturation has been shown to involve complex ne ...

Short interfering (si) RNAs are commonly used to knock down expression of proteins in mammalian cells and thereby investigate protein function. siRNAs were originally introduced into mammalian cells by transient transfection of short, synthetic, double-stranded RNA oligo nucl ...

In recent years, we have witnessed major advances in our understanding of the mammalian cell cycle and how it is regulated. Normal mammalian cellular proliferation is tightly regulated at each phase of the cell cycle by the activation and deactivation of a series of proteins that constitute the c ...

A number of different viruses interact with the cell cycle in order to subvert host-cell function and increase the efficiency of virus replication; examples can be found from DNA, retro, and RNA viruses. The majority of studies have been conducted on DNA and retroviruses whose primary site of repl ...

The method described in the following chapter utilizes a double thymidine block (an inhibitor of DNA synthesis) followed by treatment of cells with nocodazole (a mitotic inhibitor) to obtain large cell populations at distinct phases of the cell cycle. Treatment with double thymidine res ...

The determination of structures for proteins that control the eukaryotic cell cycle by nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography has made a significant contribution to our understanding of the molecular mechanisms that control cell cycle progres ...

Methods are described here to map an origin of replication in eukaryotes. Replicating DNA is enriched by BND cellulose column chromatography and by λ-exonuclease digestion; this approach has largely superceded enrichment by BrdU incorporation. The general area in which replication ...

An in situ technique for studying the chromatin binding of proteins in single fission yeast cells (Schizosaccharomyces pombe) is described. Cells are permeabilized by enzymatic digestion and extracted with a detergent-containing buffer. This procedure removes soluble protei ...