Milton Adesnik's research while affiliated with CUNY Graduate Center and other places
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Publications (34)
Publisher Summary
This chapter discusses the regulation of genes for cytochrome ρ-450. The treatment of rats with various specific drugs and xenobiotics leads to marked increases in the hepatic levels of different distinct molecular forms of cytochrome P-450 with different but overlapping substrate specificities. The broad substrate specificity of...
Insulin regulates glucose uptake into fat and muscle by modulating the distribution of the GLUT4 glucose transporter between the surface and interior of cells. The GLUT4 trafficking pathway overlaps with the general endocytic recycling pathway, but the degree and functional significance of the overlap are not known. In this study of intact adipocyt...
As a pioneer molecular cell biologist, highly skilled in both morphological and biochemical approaches, David Sabatini was a key figure in laying the foundation for the field of intracellular protein trafficking with his seminal studies on cotranslational translocation of nascent polypeptides in the endoplasmic reticulum and the intracellular sorti...
We describe an in vitro system in which post-Golgi vesicles containing metabolically labeled, sialylated, vesicular stomatitis virus (VSV) G protein molecules (VSV-G) are produced from the trans-Golgi network (TGN) of an isolated Golgi membrane fraction. This fraction is prepared from VSV-infected Madin-Darby canine kidney (MDCK) cells in which the...
We have identified and cloned the cDNA for a 912-aa protein, rab11BP, that interacts with the GTP-containing active form of rab11, a GTP-binding protein that plays a critical role in receptor recycling. Although rab11BP is primarily cytosolic, a significant fraction colocalizes with rab11 in endosomal membranes of both the sorting and recycling sub...
We identified the phosphatidylinositol transfer protein (PITP) as being responsible for a powerful latent, nucleotide-independent, Golgi-vesiculating activity that is present in the cytosol but is only manifested as an uncontrolled activity in a cytosolic protein subfraction, in which it is separated from regulatory components that appear to normal...
Rab11 is a small GTP-binding protein that in cultured mammalian cells has been shown to be concentrated in the pericentriolar endosomal recycling compartment and to play a key role in passage of the recycling transferrin receptor through that compartment [Ullrich, O., Reinsch, S., Urbé, S., Zerial, M. & Parton, R. G. (1996) J. Cell Biol. 135, 913-9...
Using a cytosol and nucleotide dependent assay that we previously developed, we have investigated the requirement for coat proteins in the in vitro production of trans-Golgi network (TGN)-derived vesicles from a Madin-Darby canine kidney (MDCK) cell Golgi fraction that contains the 35S-labeled, terminally glycosylated, envelope glycoprotein of vesi...
We have developed a system that recreates in vitro the generation of post-Golgi vesicles from an isolated Golgi fraction prepared from vesicular stomatitis virus- or influenza virus-infected Madin-Darby canine kidney or HepG2 cells. In this system, vesicle generation is temperature- and ATP-dependent and requires a supply of cytosolic proteins, inc...
The 5' flanking region of the rat PB-inducible CYP2B1 gene was isolated and the sequence from +27 to -3878 was determined. This sequence contains several putative binding sites for the liver-enriched factors HNF3 as well as an AP1, two NF-kappa B and several possible STAT sites. The promoter sequence of the CYP2B1 gene was compared to that of the C...
Cytochrome P450 (CYP) 2B1 and 2B2 are encoded by two closely related genes, CYP2B1 and CYP2B2, that are expressed at low levels in adult rat liver but are induced markedly by the administration of the drug phenobarbital (PB) or other structurally unrelated hydrophobic compounds to animals. Very little is understood about the molecular mechanisms th...
The cytochrome P450 2B subfamily in the rat contains an estimated eight to eleven members at the genomic level. Synthesis in the liver of the prototypic forms P450 2B1 and P450 2B2 is dramatically induced by phenobarbital. The 1.9-kb mRNA for P450 2B3, a third member of the P450 2B subfamily, is constitutively present in rat liver but is not induci...
The 5' flanking region of the rat PB inducible aldehyde dehydrogenase gene was isolated and the sequence from +42 to -1339 was determined. This sequence contains several putative binding sites for the liver-enriched factors HNF3 and DBP as well as a GRE and several possible AP1 sites. A TATA and CCAAT motif were assigned at positions -26 and -53. A...
NAD(P)H: quinone oxidoreductase (NQO1) is believed to be protective against cancer and toxicity caused by exposure to quinones and their metabolic precursors. This enzyme catalyzes the two-electron reduction of compounds, compared with one-electron reduction mediated by NADPH: cytochrome-P450 oxidoreductase which produces toxic and mutagenic free r...
The rat cytochrome P450 CYP2B2 gene encodes one of the two major phenobarbital-inducible forms of hepatic microsomal cytochrome P-450. The sequence of a 1.4 Kb DNA segment from the 5' flanking region of this region [Jaiswal, A., Rivkin, E. and Adesnik, M. Nucl. Acids. Res. 15: 6755 (1987)] reveals the presence of a pentadecameric oligonucleotide se...
NAD(P)H:quinone oxidoreductases (NQOs) are flavoproteins that catalyze the oxidation of NADH or NADPH by various quinones and oxidation-reduction dyes. We have previously described a complementary DNA that encodes a dioxin-inducible cytosolic form of human NAD(P)H:quinone oxidoreductase (NQO1). In the present report we describe the nucleotide seque...
A cDNA clone for rat hepatic cytochrome P450 2c (gene product IIC11) was isolated and used to study the sex specificity, expression during development, and hormonal regulation of the mRNA encoding this protein in rat liver. P450 2c mRNA levels were about 16-fold higher in males than in females and were only slightly increased in male rats after adm...
A recombinant plasmid containing a full length cDNA encoding the rat cytochrome P450IIB1 under the control of the Saccharomyces cerevisiae ADC1 promoter was constructed and transformed into the yeast strain KY118. The encoded P450IIB1 protein was produced at a level of between 0.1 and 0.2% of total yeast cellular protein (0.068 nmol/mg total cellul...
In the polymerase chain reaction (PCR) the specific amplification of a small segment of DNA within a complex DNA sample is
effected by repeated cycles of DNA denaturation and enzymatic synthesis primed by two oligonucleotides complementary to regions
within opposite strands of the DNA. In this report a simple and efficient method is described in wh...
A nomenclature for the P450 gene superfamily is proposed based on evolution. Recommendations include Roman numerals for distinct gene families, capital letters for subfamilies, and Arabic numerals for individual genes. An updating of this list, which presently includes 65 entries, will be required every 1-2 years. Assignment of orthologous genes is...
Rat hepatic cytochrome P-450 PB-1 is a prominent constitutive P-450 form whose levels increase approximately 2-3 fold upon phenobarbital administration. Antibodies raised against this protein recognized two major proteins in immunoblots of rat liver microsomal proteins and precipitated comparable amounts of two electrophoretically separable hepatic...
We have investigated the responsiveness of established rat hepatocyte cell cultures to inducers of cytochrome P-450. One Reuber hepatoma-derived line (Fu5-C8), which under normal culture conditions produces no detectable cytochrome P-450(MC) or cytochrome P-450(PB)--the major cytochrome P-450 isozymes induced by 3-methylcholanthrene and phenobarbit...
This chapter explores the application of rat hepatocyte that can serve as an excellent model for the study of the biosynthesis of endoplasmic reticulum (ER) membrane proteins. In the hepatocyte, as in all cell types, the biosynthetic machinery for the synthesis of secretory and membrane proteins includes both rough and smooth ER membrane-associated...
This chapter discusses the regulation of genes for cytochrome ρ-450. The treatment of rats with various specific drugs and xenobiotics leads to marked increases in the hepatic levels of different distinct molecular forms of cytochrome P-450 with different but overlapping substrate specificities. The broad substrate specificity of the P-450 family o...
Analysis of mRNA populations from rat liver rough microsomes and free polysomes by homologous and heterologous cDNA . mRNA hybridization shows that the two mRNA populations are distinct, demonstrating that specific mRNA classes are efficiently segregated for translation in association with endoplasmic reticulum membranes. We estimate that approxima...
The major integral membrane protein of red blood cells, the mouse equivalent of human band 3, was purified and used to raise a specfic antiserum. The murine protein resembles its human counterpart in several of its properties, including susceptibility to digestion by chymotrypsin added to intact cells and an ability to bind to concanavalin A. The s...
Treatment of Friend leukemia cells with BrdU, the thymidine analog which interferes with DMSO induced differentiation in these cells as well as the expression of differentiated character in many other cell systems, is capable of inducing erythroid differentiation. Globin mRNA, as assayed by hybridization to globin cDNA, increases 2.5− to 30−fold af...
The successful implementation of a genetic program requires that upon completion of translation polypeptides released from ribosomes are transferred to their final destination in the cell This appears to be a rather formidable process since, in a eukaryotic cell, the fate of a protein can be at least as diverse as the membranous structures within t...
Citations
... Of the five Cyp2b isoforms assessed by microarray (2b2, 2b3, 2b13, 2b15) or PCR (2b1 and 2b2) in the present study, only Cyp2b2 was clearly and strongly induced by all three triazoles. The biological functions of Cyp2b3 and Cyp2b13 have not been determined; however Cyp2b3 is not phenobarbital inducible (Jean et al., 1994). Cyp2b2 and Cyp2b15 are both induced by phenobarbital and regulated by CAR, yet only the array probe set designed for Cyp2b2 was consistently positive. ...
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... P450 family (Nebert et al., 1987). However, this system is insufficient to correlate cytochromes P450 sequences, protein structures, and function without supporting experimental evidence (Rudolf et al., 2017). ...
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... However, relatively few studies have been conducted on the differentiation inducing ability of DFMO. Differentiation of neuroblastoma cells is accompanied by an attenuation of the early peak of ODC activity and spermidine content (Chen et al., 1982a) whereas an increase in ODC acivity has been reported during differentiation of bone marrow (Rath and Reddi, 1978) and chondrocytes (Takigawa et al., 1981). Thus, cells of different origin may respond differently when treated with an inhibitor of polyamine biosynthesis . ...
... Sabatini et ai., 1975 ) and that nascent polypeptide chains contained in bound large subunits associate tightly with the underlying membranes (Adelman et al., 1973). Our recent work (Lande et al., 1975) using membrane fractions of human diploid fibroblasts, and that of Milcarek and Penman (1974) with, HeLa cells, suggest that there is also a direct association between the mRNA of bound polysomes and the ER membranes. We found that after in vitro disassembly and release of the subunits of bound polysomes mRNA nevertheless remained associated with the membranes. ...
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... In mammals, the NAD(P)H:quinone oxidoreductase (NQO) family comprises two flavin adenine dinucleotide (FAD)-dependent flavoproteins: NQO1 and NQO2 [1]. NQO1 was first identified in the rodent liver, and was designated as DT-diaphorase [2], whereas NQO2 was first isolated and characterized by screening a human liver cDNA library via hybridization with an NQO1 cDNA probe [3,4]. NQO1 and NQO2 are highly expressed in mammalian organs, with the greatest abundances detected in the liver, kidneys, and cardiovascular system [1,3,4]. ...
... In the case of MUP sites 8/9, which are conserved in at least three MUP genes (OBP3, MUP4, and LOC259246), a gene-specific amplicon [designated 8/9 (OBP3)] could be designed. STAT5 binding to all of the active MUP binding regions was male specific in liver chromatin (Fig. 6C), consistent with the strong male specificity of MUP gene expression in rat liver (47,52,53). Scatchard plot analysis for binding of STAT5 to OBP3 sites 8 and 9 (whose local sequences are also shared by sites 29 and 50 of OBP3, LOC259246, and several other MUP genes; supplemental Table S9) yielded K d values of approximately 10 -12 nM (Fig. 6D). ...
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... It is known that PB is metabolized in the liver through hydroxylation and glucuronidation mainly by CYP2C9 [46]. The administration of CBD inhibits the cytochrome P450 enzymes, an effect that results in decreased metabolism of PB [47]. Then, higher level of PB may induce toxic effects and facilitate the status epilepticus. ...
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... International Publisher pancreatic cancer [4], prostate cancer [5] and headand-neck cancer [6], express 5-to 200-fold higher levels of NQO1 than their healthy tissue counterparts. NQO1 is intimately linked with multiple carcinogenic processes [7][8][9][10]. In breast, colorectal, ovarian and cervical cancer types, elevated expression of NQO1 is closely associated with poor prognosis [3][4][5]. ...
... B, sequence of the rat CYP2B2 5 flank surrounding the GRE at 1.3 kb and the homologous region of the mouse Cyp2b10 5 flank. The two horizontal arrows indicate the two half-sites (bold) of the incomplete palindromic sequence of the putative GRE of CYP2B2 (Jaiswal et al., 1990). Under the CYP2B2 sequence are shown the differences in the Cyp2b10 homologous region. ...
... OPRK1 may influence the stress-induced reinstatement of cocaine-seeking (31,32). In a previous study, OPRK1 agonist administration was effective in decreasing cocaine-seeking and self-administration (33). ...
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