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The combination of rad mutations and regulated HO endonuclease has proven to be a potent system for elucidating DNA double-strand break (DSB) repair mechanisms in Saccharomyces cerevisiae (see Chapter 32). An analogous system comprising mutagen-sensitive mutations (see Chapter 3) ...

DNA repair is a heterogeneous process. The efficiency of repair differs not only with lesion type, but also with genomic location. Protein coding regions of the genome are repaired more efficiently than noncoding regions in a process termed preferential repair. Furthermore, for many lesi ...

Relocation into the nucleus of the yeast cytoplasmic linear plasmids was studied using a monitor plasmid pCLU1. InSaccharomyces cerevisiae, the nuclearly-relocated pCLU1 replicated in a linear form (termed pTLU-type plasmid) which carried the host telomeric repeats TG1–3 of 300– ...

Conjugated linoleic acid (CLA) can activate (in vitro) the nuclear transcription factors known as the peroxisome proliferators activated receptors (PPAR). CLA was fed at 11 g CLA/kg of feed for 45d to castrated male pigs (barrows) to better understand long term effects of PPAR activationin vi ...

The world’s oceans cover more than 70% of the earth’s surface and contain over 200,000 invertebrate and algal species (1). These organisms live in complex communities and in close association with other organisms both macro- (e.g., algae, sponges, ascidians) and micro- (e.g., nonfilamentous ba ...

The cellular retinaldehyde-binding protein (CRALBP) is expressed at high levels in vertebrate visual trssue, where it may serve to modulate the interaction of 11-cis-retinol with visual-cycle enzymes in the retinal pigment epithelium (RPE) (1). This 36-kDa protein is remarkably ster ...

Cellular retinoid-binding proteins were first identified in the mid-1970s (1). Subsequent studies established the existence of two forms of cellular retinoic acid-binding proteins (CRABPI and II) (2, 3) and two forms of cellular retinol-binding proteins (CRBP I and II) (4), each encoded by a s ...

In situ hybridization (ISH) allows the localization and relative quantification of mRNA expression for a given gene. Typically, tissue is fixed, permeablized, and then incubated with a labeled riboprobe. Once excess probe has been removed, the hybridized riboprobe that remains is dete ...

The existence of two classes of intracellular retinoid-binding proteins, the cellular retinol-binding proteins (CRBP I and II) and cellular retinoic acid-binding proteins (CRABP I and II), have been known for a long time (1). Yet, the precise roles of these proteins in retinoid physiology rem ...

Retinoids are diffusible signaling molecules important for the normal development, growth, and physiology of vertebrate organisms A role for retinoids in early embryonic development has been suggested from studies in which exogenous retinoids were administered to developing ...

Gene targeting in embryonic stem (ES) cells is a powerful technique for the modification of the mouse genome (1–3). Regarding the retinoid receptors, a number of laboratories have reported the phenotype of mice in which a given retinoic acid receptor (RAR) or retinoid X receptor (RXR) has been inac ...

An important key to understanding the function of retinoids is the determination of genes whose expression they regulate. In the past, several techniques including differential screening (1), subtractive hybridization (2), reverse transcrrptase-polymerase chain reaction ...

The electrophoretic mobility-shift assay is one of an array of techniques that are used to identify and characterize protein-DNA interactions. Thts method is based on the retardation of a labeled-DNA fragment in a nondenaturmg gel by bound proteins (1). The binding specificity and affinity ...

Steroid hormone receptors activate and repress gene transcription through an integrated series of protein-protein interactions. One principle contact is the interaction of the nuclear receptors with one another to form both homodimeric and heterodimeric complexes that bind w ...

Acute promyelocytic leukemia (APL) is one of the most common subtypes of acute myeloid leukemia (AML), accountmg for 10–15% of de novo cases and typically presenting in early middle age (1). The disease is characterized by a potentially devastating coagulopathy that can lead to rapid demise, par ...

Vitamin A (retinol), a simple fat-soluble molecule discovered in 1913 by McCollum and Davis (1), is essential for proper development, growth, and maintenance of a normal adult vertebrate organism (2–4). Dietary deficiency of vitamin A results in structural and functional abnormalities of a ...

Retinolds have been defined as a class of compounds consisting of four isoprenoid units (H2C=C(CH3)-CH=CH2) joined in a head-to-tail manner. The retinoid molecule can be divided into three parts: a trimethylated cyclohexene ring, a conjugated tetraene side chain, and a polar carbon-oxygen ...

Photoaffinity labeling and mapping of the labeled site(s) are a superior and convenient methodology for initial structural investigation of ligand-receptor complexes. The method we describe in this chapter consists of five steps.

It is widely believed that the concentrations of retinoids that activate the two classes of nuclear-retinoid receptors are crucial to the effecis of the receptors, and ultimately the actions of the retinoid-humoral system. It is therefore truly important to know the precise concentrati ...

Protein kinases (PKs) and protein phosphatases (PPases) are activated in response to extracellular stimuli such as peptidic hormones and growth factors. The resulting alteration of the phosphorylation state of target proteins is a common post-translational modification used by ...