Irina Groisman - Academia.edu (original) (raw)

Papers by Irina Groisman

Biology Direct, Jul 11, 2023

Background Prostate cancer remains one of the deadliest neoplasms in developed countries. Identif... more Background Prostate cancer remains one of the deadliest neoplasms in developed countries. Identification of new molecular markers that predict the onset and progression of the disease could improve its clinical management. Low miR-145-5p expression is consistently found in primary tumors and metastases, but the regulatory mechanisms governing its functions remain largely unknown. Methods Bioinformatics analysis was conducted to identify [1] a set of novel potential competing endogenous lncRNAs for sponging of miRNA-145-5p in prostate cancer and [2] miR-145-5p and other EMT-related miRNAs response elements in lnc-ZNF30-3. Quantification of miR-145-5p, lnc-ZNF30-3, and TWIST1 expression levels in tumor tissues in RNA sequencing datasets of our and TCGA PRAD cohorts revealed a correlation with clinical outcome of prostate cancer patients. Biochemical and cell biology approaches, such as RNA pull-down, western blot, immunostaining, and wound healing assays were used for evaluation of the impact of TWIST1/miR-145/ lnc-ZNF30-3 interactions in prostate cancer cells altered in miRNA and lncRNA expression. Results We identified a few potential lncRNA sponges of miR-145-5p, including lnc-ZNF30-3. It contains five response elements for miR-145-5p, but also other miRNAs targeting EMT transcription factors. Lnc-ZNF30-3 is significantly upregulated in prostate cancer cell lines and tumor tissues, and its high expression is correlated with poor patient prognosis. We demonstrated that lnc-ZNF30-3 is associated with AGO2 and specifically interacts with the miR-145-5p seed region. Knockdown of lnc-ZNF30-3 results in decreased migration of prostate cancer cells and downregulation of EMT drivers such as TWIST1 and ZEB1 at both the RNA and protein levels. These phenotypic and molecular features of lnc-ZNF30-3-depleted cells are partially rescued by miR-145-5p inhibition. Conclusions Collectively, our results point to lnc-ZNF30-3 as a novel competing endogenous lncRNA for miR-145-5p and other miRNAs that target TWIST1 as well as other EMT transcription factors. Prostate cancer patients with

Scientific Reports, May 1, 2016

The cytoplasmic element binding protein 1 (CPEB1) regulates many important biological processes r... more The cytoplasmic element binding protein 1 (CPEB1) regulates many important biological processes ranging from cell cycle control to learning and memory formation, by controlling mRNA translation efficiency via 3′ untranslated regions (3′UTR). In the present study, we show that CPEB1 is significantly downregulated in human Glioblastoma Multiforme (GBM) tissues and that the restoration of its expression impairs glioma cell lines growth. We demonstrate that CPEB1 promotes the expression of the cell cycle inhibitor p27 Kip1 by specifically targeting its 3′UTR, and competes with miR-221/222 binding at an overlapping site in the 3′UTR, thus impairing miR-221/222 inhibitory activity. Upon binding to p27 Kip1 3′UTR, CPEB1 promotes elongation of poly-A tail and the subsequent translation of p27 Kip1 mRNA. This leads to higher levels of p27 Kip1 in the cell, in turn significantly inhibiting cell proliferation, and confers to CPEB1 a potential value as a tumor suppressor in Glioblastoma. The cytoplasmic element binding protein 1 (CPEB1) is the founding member of a family of four conserved sequence-specific RNA-binding proteins (CPEB1-4) that regulate gene expression at the post-transcriptional level. 1 CPEBs specifically bind to the cytoplasmic polyadenylation element (CPE; consensus sequence UUUUUAU) in 3′ UTRs of messenger RNAs and control, along with other cellular factors, both translational repression and activation through regulation of poly(A) tail length 2. Initially CPEBs were identified as maternal mRNA regulators, responsible of cytoplasmic polyadenylation of dormant mRNAs with short poly(A) tails and consequent induction of their translation during early embryonic development 3,4. In the last years, the data about the biochemistry and biology of CPEBs has grown rather substantially and it is now clear that these proteins are key mediators of several important biological processes ranging from cell cycle control and cancer to learning and memory 5,6. Some evidences suggest a potential tumor suppressive role for CPEB1. The expression level of CPEB1 mRNA were found decreased in ovarian, melanoma and gastric cancer as well as in cell lines derived from breast, myeloma and colorectal cancer 7-11. This reduction has been associated with the capacity of malignant cells to promote invasion, angiogenesis, to increase resistance to nutritional stress and to induce epithelial-to-mesenchymal transition 12,13. Moreover, CPEB1-knockout human foreskin fibroblasts proliferate more rapidly, bypassing senescence and are subject to the "Warburg effect", an aerobic glycolysis considered to be a hallmark of cancer cells 14. GBM is the most common and aggressive primary brain tumor. Despite interventional therapy, the overall prognosis for GBM patients remains poor 15 ; thus, it is essential to understand its molecular pathogenesis, in order to provide new insight into modern therapy. Among factors whose mis-regulation has been linked to GBM progression, the cyclin-dependent kinase inhibitor p27 Kip1 is an established prognostic marker and its expression has been inversely related to tumor grade and positively related to favorable outcome of GBM patients 16,17. p27 Kip1 plays a pivotal role in the control of cell proliferation, differentiation, and apoptosis 17-19 ; it is considered

Cell, May 1, 2002

Translational Control of the Embryonic Cell Cycle which in many respects are similar to the mitot... more Translational Control of the Embryonic Cell Cycle which in many respects are similar to the mitotic divi-Irina Groisman, Mi-Young Jung, sions of somatic cells (Sheets et al., 1994; Stebbins-Madathia Sarkissian, Quiping Cao, Boaz et al., 1996). Oocytes are arrested at the end of and Joel D. Richter 1 prophase I, which resembles G2 of the somatic cell Program in Molecular Medicine cycle. When stimulated by progesterone, the oocytes University of Massachusetts Medical School synthesize cyclin B1, form active MPF, reenter the mei-Worcester, Massachusetts 01605 otic divisions (oocyte maturation), and progress to metaphase I (MI) (Swenson et al., 1986). A subsequent decrease in MPF activity is necessary for progression to Summary anaphase I (AI), while a second increase then drives the oocytes to metaphase II (Nebreda and Ferby, 2000). The synthesis and destruction of cyclin B drives mito-During meiosis, the translational control of cyclin B1 sis in eukaryotic cells. Cell cycle progression is also and other mRNAs such as mos, cyclin A1, and cdk1 is regulated at the level of cyclin B translation. In cycling regulated by cytoplasmic polyadenylation (Sheets et al., extracts from Xenopus embryos, progression into M 1995; Stebbins-Boaz et al., 1996; de Moor and Richter, phase requires the polyadenylation-induced transla-1997; reviewed by Richter, 2000; Wickens et al., 2000; tion of cyclin B1 mRNA. Polyadenylation is mediated Mendez and Richter, 2001). When dormant in oocytes, by the phosphorylation of CPEB by Aurora, a kinase these mRNAs contain short poly(A) tails, and it is only whose activity oscillates with the cell cycle. Exit from when the tails are elongated during maturation that M phase seems to require deadenylation and subsetranslation ensues. Polyadenylation in turn is regulated quent translational silencing of cyclin B1 mRNA by by two cis-acting sequences in the 3Ј untranslated re-Maskin, a CPEB and eIF4E binding factor, whose exgions (UTRs) of responding mRNAs, the cytoplasmic pression is cell cycle regulated. These observations polyadenylation element (CPE), which has the general suggest that regulated cyclin B1 mRNA translation is structure of UUUUUAU, and the hexanucleotide AAU essential for the embryonic cell cycle. Mammalian AAA (McGrew et al., 1989; Fox et al., 1989). CPEB, an cells also display a cell cycle-dependent cytoplasmic RNA recognition motif (RRM) and zinc finger-containing polyadenylation, suggesting that translational control protein, binds the CPE (Hake and Richter, 1994; Hake et by polyadenylation might be a general feature of mitoal., 1998), while cleavage and polyadenylation specificity sis in animal cells. factor (CPSF) interacts with the AAUAAA (Dickson et al., 1999). Polyadenylation is initiated by the phosphoryla

Genes & Development, 2006

... Control of cellular senescene by CPEB. Autores: Irina Groisman, Veronica Martin, Roger J. Dav... more ... Control of cellular senescene by CPEB. Autores: Irina Groisman, Veronica Martin, Roger J. Davis, Norman J. Kennedy, Maria Ivshina, Joel D. Richter; Localización: Genes & development, ISSN 0890-9369, Nº. 19, 2006 , pág. 7. Fundación Dialnet. Acceso de usuarios registrados ...

World Journal of Biological Chemistry, 2014

circumstances, such as their expression levels, affinity to the binding sites, and localization i... more circumstances, such as their expression levels, affinity to the binding sites, and localization in the cell, which can be controlled by various physiological conditions. Moreover, the functional and/or physical interactions of the factors binding to 3'UTR can change the character of their actions. These interactions vary during the cell cycle and in response to changing physiological conditions. Abnormal functioning of the factors can lead to disease. In this review we will discuss how alterations of these factors or their interaction can affect cancer development and promote or enhance the malignant phenotype of cancer cells. Understanding these alterations and their impact on 3'UTR-directed posttranscriptional gene regulation will uncover promising new targets for therapeutic intervention and diagnostics. We will also discuss emerging new tools in cancer diagnostics and therapy based on 3'UTR binding factors and approaches to improve them.

Biochimie, May 1, 1990

A positive correlation between poly(U) misreading and efficiency of poly(dT) translation has been... more A positive correlation between poly(U) misreading and efficiency of poly(dT) translation has been revealed in cell-free systems from wild-type E coli and streptomycin--resistant mutants with altered ribosomal protein S12. Different factors promoting misreading of poly(U) such as aminoglycoside antibiotics and Mg2+ ions also stimulate poly(dT) translation. The effect of the antibiotics on poly(U) translation efficiency and misreading as well as on poly(dT) decoding is characterised by the same order: neomycin greater than kanamycin greater than streptomycin. S12 mutants ribosomes are less erroneous in poly(U) translation and less efficient in poly(dT) decoding. The data obtained are in good agreement with the hypothesis of stereospecific stabilization of codon-anticodon complexes by the ribosome decoding centre.

Cold Spring Harbor Symposia on Quantitative Biology, 2001

... EMBO J. 17: 1053. de Moor CH and Richter JD 1997. The Mos pathway regulates cytoplasmic polya... more ... EMBO J. 17: 1053. de Moor CH and Richter JD 1997. The Mos pathway regulates cytoplasmic polyadenylation in Xenopus oocytes. Mol. Cell. Biol. 17: 6419. . ... Annu. Rev. Biochem. 68: 913. Groisman I., Huang YS, Mendez R., Cao Q., Therukauf W., and Richter JD 2000. ...

Genes & Development, Oct 1, 2006

Cytoplasmic polyadenylation element-binding protein (CPEB) is a sequence-specific RNA-binding pro... more Cytoplasmic polyadenylation element-binding protein (CPEB) is a sequence-specific RNA-binding protein that promotes polyadenylation-induced translation. While a CPEB knockout (KO) mouse is sterile but overtly normal, embryo fibroblasts derived from this mouse (MEFs) do not enter senescence in culture as do wild-type MEFs, but instead are immortal. Exogenous CPEB restores senescence in the KO MEFs and also induces precocious senescence in wild-type MEFs. CPEB cannot stimulate senescence in MEFs lacking the tumor suppressors p53, p19 ARF , or p16 INK4A ; however, the mRNAs encoding these proteins are unlikely targets of CPEB since their expression is the same in wild-type and KO MEFs. Conversely, Ras cannot induce senescence in MEFs lacking CPEB, suggesting that it may lie upstream of CPEB. One target of CPEB regulation is myc mRNA, whose unregulated translation in the KO MEFs may cause them to bypass senescence. Thus, CPEB appears to act as a translational repressor protein to control myc translation and resulting cellular senescence.

Cell Cycle, Mar 3, 2016

MicroRNAs (miRNAs) in the AGO-containing RISC complex control messenger RNA (mRNA) translation by... more MicroRNAs (miRNAs) in the AGO-containing RISC complex control messenger RNA (mRNA) translation by binding to mRNA 3 0 untranslated region (3 0 UTR). The relationship between miRNAs and other regulatory factors that also bind to mRNA 3 0 UTR, such as CPEB1 (cytoplasmic polyadenylation element-binding protein), remains elusive. We found that both CPEB1 and miR-15b control the expression of WEE1, a key mammalian cell cycle regulator. Together, they repress WEE1 protein expression during G1 and S-phase. Interestingly, the 2 factors lose their inhibitory activity at the G2/M transition, at the time of the cell cycle when WEE1 expression is maximal, and, moreover, rather activate WEE1 translation in a synergistic manner. Our data show that translational regulation by RISC and CPEB1 is essential in cell cycle control and, most importantly, is coordinated, and can be switched from inhibition to activation during the cell cycle.

Biopolymers & Cell, Jan 20, 2013

Eukaryotic translation elongation factor 1A exists as two 98 % homologous isoforms: eEF1A1 (A1) a... more Eukaryotic translation elongation factor 1A exists as two 98 % homologous isoforms: eEF1A1 (A1) and eEF1A2 (A2) which are tissue and development specific. Despite high homology in an open reading frame (ORF) region, mRNAs coding for eEF1A1 and eEF1A2 are different in their untranslated regions (UTR), suggesting a possibility of their dissimilar post-transcriptional regulation. Aim. To analyze the existence of cis-acting motifs in the UTRs of EEF1A1/A2 mRNAs, to confirm the possibility of post-transcriptional control of eEF1A1 and eEF1A2 expression. Methods. An ensemble of bioinformatic methods was applied to predict regulatory motifs in the UTRs of EEF1A1/A2 mRNAs. Dual-luciferase reporter assay was employed to detect post-transcriptional regulation of eEF1A1/A2 expression. Results. Numerous regulatory motifs in the UTR of EEF1A1/A2 mRNAs were found bioinformatically. The experimental evidence was obtained for the existence of negative regulation of EEF1A1 and positive regulation of EEF1A2 mRNA in the model of breast cancer development. Conclusions. EEF1A1 and EEF1A2 mRNAs contain distinct motifs in the UTRs and are differently regulated in cancer suggesting the possibility of their control by different cellular signals.

Biopolymers & Cell, Nov 20, 2012

British Journal of Cancer, May 21, 2013

Background: Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-oncogene. We p... more Background: Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-oncogene. We proposed that stimulation of the eEF1A2 expression in cancer tissues is caused by the loss of miRNA-mediated control. Methods: Impact of miRNAs on eEF1A2 at the mRNA and protein levels was examined by qPCR and western blot, respectively. Dual-luciferase assay was applied to examine the influence of miRNAs on 3 0-UTR of EEF1A2. To detect miRNA-binding sites, mutations into the 3 0-UTR of EEF1A2 mRNA were introduced by the overlap extension PCR. Results: miR-663 and miR-744 inhibited the expression of luciferase gene attached to the 3 0-UTR of EEF1A2 up to 20% and 50%, respectively. In MCF7 cells, overexpression of miR-663 and miR-744 reduced the EEF1A2 mRNA level by 30% and 50%. Analogous effects were also observed at the eEF1A2 protein level. In resveratrol-treated MCF7 cells the upregulation of mir-663 and mir-744 was accompanied by downregulation of EEF1A2 mRNA. Both miRNAs were able to inhibit the proliferation of MCF7 cells. Conclusion: miR-663 and miR-744 mediate inhibition of the proto-oncogene eEF1A2 expression that results in retardation of the MCF7 cancer cells proliferation. Antitumour effect of resveratrol may include stimulation of the miR-663 and miR-744 expression. Eukaryotic translation elongation factor 1A (eEF1A), one of the key players in protein biosynthesis, binds aminoacyl-tRNA and transfers it to the A-site of the ribosome. Human eEF1A exists as two isoforms: eEF1A1, its gene located on 6q13, and eEF1A2, positioned on 20q13 (Knudsen et al, 1993). The two proteins are 92% identical and 98% similar. Both proteins are tissue-and development-specific. Eukaryotic translation elongation factor 1A2 performs nascent polypeptide elongation on the 80S ribosome in neuronal, muscle and cardiac tissues, being the only representative of the elongation factor 1 family (Lee et al, 1992). Other tissues of higher vertebrates employ eEF1A1 for this purpose. Developmentally, the A1 isoform is replaced by A2 in muscles and neurons during the early postnatal period (Pan et al, 2004). Owing to the mutually exclusive tissue-specific character of their expression, the isoforms are rarely found expressed together in the tissues under

Oncogene, Jan 23, 2012

TWIST1 is a highly conserved basic helix-loop-helix transcription factor that promotes epithelial... more TWIST1 is a highly conserved basic helix-loop-helix transcription factor that promotes epithelial-mesenchymal transition (EMT). Its misregulation has been observed in various types of tumors. Using the MCF-10A-series of cell lines that recapitulate the early stages of breast cancer formation and EMT, we found TWIST1 to be upregulated during EMT and downregulated early in carcinogenesis. The TWIST1 3 0 UTR contains putative regulatory elements, including miRNA target sites and two cytoplasmic polyadenylation elements (CPE). We found that miR-580, CPEB1, and CPEB2 act as negative regulators of TWIST1 expression in a sequence-specific and additive/cooperative manner.

Cell, Oct 1, 2000

quences in the 3Ј untranslated regions (UTRs) of responding mRNAs are required for cytoplasmic po... more quences in the 3Ј untranslated regions (UTRs) of responding mRNAs are required for cytoplasmic polyadenylation: the hexanucleotide AAUAAA and the cytoplasmic element (CPE), whose general structure is UUUUUAU (McGrew et al., 1989; Fox et al., 1989

Biology Direct

Biochimie, 1990

Biopolymers and Cell, 1989

British journal of cancer, Jan 11, 2013

Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-oncogene. We proposed that... more Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-oncogene. We proposed that stimulation of the eEF1A2 expression in cancer tissues is caused by the loss of miRNA-mediated control. Impact of miRNAs on eEF1A2 at the mRNA and protein levels was examined by qPCR and western blot, respectively. Dual-luciferase assay was applied to examine the influence of miRNAs on 3'-UTR of EEF1A2. To detect miRNA-binding sites, mutations into the 3'-UTR of EEF1A2 mRNA were introduced by the overlap extension PCR. miR-663 and miR-744 inhibited the expression of luciferase gene attached to the 3'-UTR of EEF1A2 up to 20% and 50%, respectively. In MCF7 cells, overexpression of miR-663 and miR-744 reduced the EEF1A2 mRNA level by 30% and 50%. Analogous effects were also observed at the eEF1A2 protein level. In resveratrol-treated MCF7 cells the upregulation of mir-663 and mir-744 was accompanied by downregulation of EEF1A2 mRNA. Both miRNAs were able to inhibit the prolifer...

Oncogene, 2012