Antizyme Affects Cell Proliferation and Viability Solely through Regulating Cellular Polyamines

Bercovich, Z.; Snapir, Zohar; Keren-Paz, Alona; Kahana, Chaim

doi:10.1074/jbc.m111.270637

Cited by 24 publications

(21 citation statements)

References 34 publications

(42 reference statements)

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“…The identification of OAZ1 as a gene whose silencing can enhance recombinant protein production is an indication that this gene normally suppresses protein synthesis. This is compatible with the known function of the OAZ1 as a negative regulator of polyamine homeostasis, cell proliferation, and transformation (Bercovich et al, ; Coffino, ; Kahana, ; Pegg, ). OAZ1 is a negative regulator of ODC, a rate‐limiting enzyme in polyamine biosynthesis.…”

Section: Discussionsupporting

confidence: 84%

Genome‐scale RNA interference screen identifies antizyme 1 (OAZ1) as a target for improvement of recombinant protein production in mammalian cells

Xiao

Chen

Buehler

et al. 2016

Biotech & Bioengineering

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For the purpose of improving recombinant protein production from mammalian cells, an unbiased, high-throughput whole-genome RNA interference screen was conducted using human embryonic kidney 293 (HEK 293) cells expressing firefly luciferase. 21,585 human genes were individually silenced with three different siRNAs for each gene. The screen identified 56 genes that led to the greatest improvement in luciferase expression. These genes were found to be included in several pathways involved in spliceosome formation and mRNA processing, transcription, metabolic processes, transport and protein folding. The 10 genes that most enhanced protein expression when down regulated, were further confirmed by measuring the effect of their silencing on the expression of three additional recombinant proteins. Among the confirmed genes, OAZ1- the gene encoding the ornithine decarboxylase antizyme1- was selected for detailed investigation, since its silencing improved the reporter protein production without affecting cell viability. Silencing OAZ1 caused an increase of the ornithine decarboxylase enzyme and the cellular levels of putrescine and spermidine; an indication that increased cellular polyamines enhances luciferase expression without affecting its transcription. The study shows that OAZ1 is a novel target for improving expression of recombinant proteins. The genome-scale screening performed in this work can establish the foundation for targeted design of an efficient mammalian cell platform for various biotechnological applications.

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Section: Discussionsupporting

confidence: 84%

Genome‐scale RNA interference screen identifies antizyme 1 (OAZ1) as a target for improvement of recombinant protein production in mammalian cells

Xiao

Chen

Buehler

et al. 2016

Biotech & Bioengineering

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“…In addition, it is also likely that AZIN2 may affect the degradation of antizyme-targeted proteins different to ODC, since the involvement of AZ1 in the degradation of cell cycle related proteins has been reported [35], [36]. Although this later claim has been recently questioned [37], it does not rule out the possibility that AZIN2, either directly or by acting on antizymes, could regulate proteins implicated in secretory processes. In this novel scenario, AZIN2 would affect cellular processes through its action on target proteins in addition to modulate intracellular polyamine pools.…”

Section: Discussionmentioning

confidence: 99%

Antizyme Inhibitor 2 Hypomorphic Mice. New Patterns of Expression in Pancreas and Adrenal Glands Suggest a Role in Secretory Processes

et al. 2013

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The intracellular levels of polyamines, polycations implicated in proliferation, differentiation and cell survival, are regulated by controlling their biosynthesis, catabolism and transport. Antizymes and antizyme inhibitors are key regulatory proteins of polyamine levels by affecting ornithine decarboxylase, the rate-limiting biosynthetic enzyme, and polyamine uptake. We recently described the molecular function of a novel antizyme inhibitor (AZIN2). However, the physiological function of AZIN2 in mammals is mostly unknown. To gain insight on the tissue expression profile of AZIN2 and to find its possible physiological role, we have generated, transgenic mice with severe Azin2 hypomorphism. This mouse model expresses transgenic bacterial β-D-galactosidase as a reporter gene, under the control of the Azin2 endogenous promoter, what allows a very sensitive and specific detection of the expression of the gene in the different tissues of transgenic mice. The biochemical and histochemical analyses of β-D-galactosidase together with the quantification of Azin2 mRNA levels, corroborated that AZIN2 is mainly expressed in testis and brain, and showed for the first time that AZIN2 is also expressed in the adrenal glands and pancreas. In these tissues, AZIN2 was not expressed in all type of cells, but rather in specific type of cells. Thus, AZIN2 was mainly found in the haploid germinal cells of the testis and in different brain regions such as hippocampus and cerebellum, particularly in specific type of neurons. In the adrenal glands and pancreas, the expression was restricted to the adrenal medulla and to the Langerhans islets, respectively. Interestingly, plasma insulin levels were significantly reduced in the transgenic mice. These results support the idea that AZIN2 may have a role in the modulation of reproductory and secretory functions and that this mouse model might be an interesting tool for the progress of our understanding on the role of AZIN2 and polyamines in specific mammalian cells.

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“…The polyamines (putrescine, spermidine, and spermine) are present in all mammalian cells (Zwierzchowski et al, 1986), and participate in multiple cellular processes, such as cell proliferation (Bercovich et al, 2011), apoptosis (Tome et al, 1997), signal transduction (Bachrach et al, 2001;Pignatti et al, 2004), amino acid metabolism (Lef evre et al, 2011b), oxidative processes (Smirnova et al, 2012), and DNA and histone methylation (Brooks, 2012;Karouzakis et al, 2012). All of these processes are important for the development of the endometrial glands and their secretory activity.…”

Section: Introductionmentioning

confidence: 99%

Regulation of the polyamine metabolic pathway in the endometrium of cows during early diestrus

Ramos

Mesquita

D’Alexandri

et al. 2014

Molecular Reproduction Devel

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The timing and magnitude of exposure to preovulatory estradiol followed by post-ovulatory progesterone (periovulatory endocrine milieu) in cattle modulate endometrial gene expression, histotroph composition, and conceptus development, but the mechanisms underlying this regulation remain unknown. Using an experimental model based on the modulation of follicle growth, this work aimed to evaluate if the polyamine metabolic pathway is regulated by the periovulatory endocrine milieu. Nelore cows were manipulated to ovulate small (n = 15) or large (n = 15) follicles, then the profiles of polyamines and their synthetic enzymes were compared between groups. Transcripts for the enzymes of this pathway, ornithine decarboxylase 1 (ODC1; the rate-limiting enzyme in polyamine biosynthesis) protein quantification, adenosylmethionine decarboxylase 1 (AMD1) protein immunolocalization, and concentrations of the different polyamines (putrescine, spermidine, and spermine) were respectively quantified by quantitative reverse-transcriptase PCR, immunoblotting, immunohistochemistry, and gas chromatography-mass spectrometry in both the endometrium and uterine flushing. No differences in gene and protein expression or concentration of polyamines were observed between groups. There were significant correlations between the relative abundance of ODC1 and spermidine/spermine N1-acetyltransferase 1 (SAT1) transcripts as well as between antizyme inhibitor 1 (AZIN1) and adenosylmethionine decarboxylase 1 (AMD1) transcripts. In conclusion, our results show that the polyamine metabolic pathway is present and functional, but not regulated by the periovulatory endocrine milieu in the bovine endometrium.

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Antizyme Affects Cell Proliferation and Viability Solely through Regulating Cellular Polyamines

Cited by 24 publications

References 34 publications

Genome‐scale RNA interference screen identifies antizyme 1 (OAZ1) as a target for improvement of recombinant protein production in mammalian cells

Genome‐scale RNA interference screen identifies antizyme 1 (OAZ1) as a target for improvement of recombinant protein production in mammalian cells

Antizyme Inhibitor 2 Hypomorphic Mice. New Patterns of Expression in Pancreas and Adrenal Glands Suggest a Role in Secretory Processes

Regulation of the polyamine metabolic pathway in the endometrium of cows during early diestrus

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