TY  - JOUR
AU  - Lazić, Andrijana
AU  - Ferrai, Carmelo
AU  - Stucchi, Laura
AU  - Prada, Ilaria
AU  - Podini, Paola
AU  - Baba, Tadashi
AU  - Rocchi, Mariano
AU  - Meldolesi, Jacopo
AU  - D'Alessandro, Rosalba
PY  - 2009
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/350
AB  - Expression of neurosecretion by nerve cells requires the levels of the transcription repressor element-1 silencing transcription factor (REST) to be very low. However, when high-REST clones of PC12 cells, defective of neurosecretion, were fused to other high-REST, non-neurosecretory cells, some neurosecretion was recovered. To clarify the mechanism of this recovery, we fused defective PC12 cells with human lymphocytes. A cytogenetic analysis revealed all hybrid clones that recovered neurosecretion to contain a fragment of chromosome 11 including the gene encoding BHC80, a protein of one of the complexes that mediate REST repression. In these clones, REST levels were as high as in defective PC12, whereas BHC80, localized in the nucleus, was 4- to 5-fold higher. Transient transfection of defective PC12 with various amounts of BHC80 cDNA induced (1) in defective PC12, the reexpression of only neurosecretion mRNAs; (2) in defective PC12 cotransfected with the REST negative construct DNA-binding domain (to attenuate gene repression), the recovery of a weak, but complete neurosecretory phenotype, including dense-core granules and their regulated exocytosis. Chromatin immunoprecipitation and immunodepletion analyses revealed the extensive BHC80 association with REST at the genes of two neurosecretion proteins, chromograninB and SNAP25, however only in the low-REST PC12, whereas in high-REST defective PC12 no association was appreciable. In defective PC12 transfected with BHC80 some association was reestablished. Therefore, the recovery of neurosecretion observed after fusion/transfection of defective PC12 depends on the reciprocal level of BHC80 and REST, with BHC80 working as a negative modulator of REST repression. This role appears of possible cell physiological and pathological importance.
PB  - Soc Neuroscience, Washington
T2  - Journal of Neuroscience
T1  - The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex
EP  - 6307
IS  - 19
SP  - 6296
VL  - 29
DO  - 10.1523/JNEUROSCI.5943-08.2009
ER  - 

@article{
author = "Lazić, Andrijana and Ferrai, Carmelo and Stucchi, Laura and Prada, Ilaria and Podini, Paola and Baba, Tadashi and Rocchi, Mariano and Meldolesi, Jacopo and D'Alessandro, Rosalba",
year = "2009",
abstract = "Expression of neurosecretion by nerve cells requires the levels of the transcription repressor element-1 silencing transcription factor (REST) to be very low. However, when high-REST clones of PC12 cells, defective of neurosecretion, were fused to other high-REST, non-neurosecretory cells, some neurosecretion was recovered. To clarify the mechanism of this recovery, we fused defective PC12 cells with human lymphocytes. A cytogenetic analysis revealed all hybrid clones that recovered neurosecretion to contain a fragment of chromosome 11 including the gene encoding BHC80, a protein of one of the complexes that mediate REST repression. In these clones, REST levels were as high as in defective PC12, whereas BHC80, localized in the nucleus, was 4- to 5-fold higher. Transient transfection of defective PC12 with various amounts of BHC80 cDNA induced (1) in defective PC12, the reexpression of only neurosecretion mRNAs; (2) in defective PC12 cotransfected with the REST negative construct DNA-binding domain (to attenuate gene repression), the recovery of a weak, but complete neurosecretory phenotype, including dense-core granules and their regulated exocytosis. Chromatin immunoprecipitation and immunodepletion analyses revealed the extensive BHC80 association with REST at the genes of two neurosecretion proteins, chromograninB and SNAP25, however only in the low-REST PC12, whereas in high-REST defective PC12 no association was appreciable. In defective PC12 transfected with BHC80 some association was reestablished. Therefore, the recovery of neurosecretion observed after fusion/transfection of defective PC12 depends on the reciprocal level of BHC80 and REST, with BHC80 working as a negative modulator of REST repression. This role appears of possible cell physiological and pathological importance.",
publisher = "Soc Neuroscience, Washington",
journal = "Journal of Neuroscience",
title = "The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex",
pages = "6307-6296",
number = "19",
volume = "29",
doi = "10.1523/JNEUROSCI.5943-08.2009"
}

Lazić, A., Ferrai, C., Stucchi, L., Prada, I., Podini, P., Baba, T., Rocchi, M., Meldolesi, J.,& D'Alessandro, R.. (2009). The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex. in Journal of Neuroscience
Soc Neuroscience, Washington., 29(19), 6296-6307.
https://doi.org/10.1523/JNEUROSCI.5943-08.2009

Lazić A, Ferrai C, Stucchi L, Prada I, Podini P, Baba T, Rocchi M, Meldolesi J, D'Alessandro R. The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex. in Journal of Neuroscience. 2009;29(19):6296-6307.
doi:10.1523/JNEUROSCI.5943-08.2009 .

Lazić, Andrijana, Ferrai, Carmelo, Stucchi, Laura, Prada, Ilaria, Podini, Paola, Baba, Tadashi, Rocchi, Mariano, Meldolesi, Jacopo, D'Alessandro, Rosalba, "The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex" in Journal of Neuroscience, 29, no. 19 (2009):6296-6307,
https://doi.org/10.1523/JNEUROSCI.5943-08.2009 . .

TY  - JOUR
AU  - D'Alessandro, Rosalba
AU  - Lazić, Andrijana
AU  - Stucchi, Laura
AU  - Podini, Paola
AU  - Malosio, Maria Luisa
AU  - Meldolesi, Jacopo
PY  - 2008
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/336
AB  - The neurosecretory process is acquired during differentiation and can be lost en block by differentiated cells. To investigate the role of REST/NRSF, a transcription repressor, in the maintenance of the process we studied two PC12 clones, one wt and one defective, expressing low and high levels of endogenous RE-1 silencing transcription (factor) (REST), respectively. Stable transfection of constructs demonstrated that REST represses 10 genes coding for proteins of neurosecretory vesicles and their exocytosis, eight including and two lacking the REST-binding sequence, RE-1. Of these genes, those of chromogranins were strongly repressed by fewfold increases of REST, those of VAMP2 and syntaxin1a required much higher levels. Moreover, in wt cells transfected with an active construct the dense-core vesicles, still competent for regulated exocytosis, were much smaller, with lighter cores; in defective cells, the dominant-negative construct induced the rescue of many vesicle/exocytosis genes but not of those of chromogranins. Small dense-core vesicles, exocytized upon stimulation, were rescued when the construct-transfected defective cells were transfected also with chromograninA or treated with trichostatinA, a blocker of histone deacetylases. Our results identify REST, working by direct and indirect mechanisms, as the factor governing the maintenance of the neurosecretory process and the properties of dense-core vesicles in PC12 cells.
PB  - Blackwell Publishing, Oxford
T2  - Journal of Neurochemistry
T1  - Expression of the neurosecretory process in pc12 cells is governed by rest
EP  - 1383
IS  - 4
SP  - 1369
VL  - 105
DO  - 10.1111/j.1471-4159.2008.05259.x
ER  - 

@article{
author = "D'Alessandro, Rosalba and Lazić, Andrijana and Stucchi, Laura and Podini, Paola and Malosio, Maria Luisa and Meldolesi, Jacopo",
year = "2008",
abstract = "The neurosecretory process is acquired during differentiation and can be lost en block by differentiated cells. To investigate the role of REST/NRSF, a transcription repressor, in the maintenance of the process we studied two PC12 clones, one wt and one defective, expressing low and high levels of endogenous RE-1 silencing transcription (factor) (REST), respectively. Stable transfection of constructs demonstrated that REST represses 10 genes coding for proteins of neurosecretory vesicles and their exocytosis, eight including and two lacking the REST-binding sequence, RE-1. Of these genes, those of chromogranins were strongly repressed by fewfold increases of REST, those of VAMP2 and syntaxin1a required much higher levels. Moreover, in wt cells transfected with an active construct the dense-core vesicles, still competent for regulated exocytosis, were much smaller, with lighter cores; in defective cells, the dominant-negative construct induced the rescue of many vesicle/exocytosis genes but not of those of chromogranins. Small dense-core vesicles, exocytized upon stimulation, were rescued when the construct-transfected defective cells were transfected also with chromograninA or treated with trichostatinA, a blocker of histone deacetylases. Our results identify REST, working by direct and indirect mechanisms, as the factor governing the maintenance of the neurosecretory process and the properties of dense-core vesicles in PC12 cells.",
publisher = "Blackwell Publishing, Oxford",
journal = "Journal of Neurochemistry",
title = "Expression of the neurosecretory process in pc12 cells is governed by rest",
pages = "1383-1369",
number = "4",
volume = "105",
doi = "10.1111/j.1471-4159.2008.05259.x"
}

D'Alessandro, R., Lazić, A., Stucchi, L., Podini, P., Malosio, M. L.,& Meldolesi, J.. (2008). Expression of the neurosecretory process in pc12 cells is governed by rest. in Journal of Neurochemistry
Blackwell Publishing, Oxford., 105(4), 1369-1383.
https://doi.org/10.1111/j.1471-4159.2008.05259.x

D'Alessandro R, Lazić A, Stucchi L, Podini P, Malosio ML, Meldolesi J. Expression of the neurosecretory process in pc12 cells is governed by rest. in Journal of Neurochemistry. 2008;105(4):1369-1383.
doi:10.1111/j.1471-4159.2008.05259.x .

D'Alessandro, Rosalba, Lazić, Andrijana, Stucchi, Laura, Podini, Paola, Malosio, Maria Luisa, Meldolesi, Jacopo, "Expression of the neurosecretory process in pc12 cells is governed by rest" in Journal of Neurochemistry, 105, no. 4 (2008):1369-1383,
https://doi.org/10.1111/j.1471-4159.2008.05259.x . .