Raúl Aguilar-Roblero was born in Tuxtla Gutierrez, Chiapas in México in 1959. He attended the Universidad Nacional Autónoma de México, from 1975 to 1989, where he graduated as B.Sc. M.Sc., M.D., Ph.D. He married Beatríz Eugenia Gutierrez-Ruiz in 1982 until her death form Ovaric Cancer in 2020. They had two children Raúl (1984) and Eugenia (1988). Supervisors: Dr. Rene Drucker-Colin and Robert Yates Moore
... DOI: 10.1080/09291010400028542 Rebeca Toledo a c , Raúl Aguilar-Roblero b , Enrique Canchola ... more ... DOI: 10.1080/09291010400028542 Rebeca Toledo a c , Raúl Aguilar-Roblero b , Enrique Canchola c & Mario Caba Dr a * pages 47-55. Available online: 02 Sep 2011. ...
A characteristic mechanism of gene expression regulation during seed germination is the selective... more A characteristic mechanism of gene expression regulation during seed germination is the selective translation of mRNAs. Previous findings indicate that the two cap-binding complexes eIF4F (with eIF4E and eIF4G subunits) and eIF(iso)4F [with eIF(iso)4E and eIF(iso)4G subunits] are differentially expressed during maize seed germination. In addition, several studies in vitro have suggested that these factors may participate in selective mRNA translation. The translational activities of eIF4E and eIF(iso)4E were tested in vitro using transcripts from two different sets: dry (0 h) and 24-h-imbibed maize embryonic axes. In vitro translation of these mRNA pools in the presence of the recombinant eIF4E or eIF(iso)4E, and the native cap-binding complexes from dry- or 24-h-imbibed axes, produced different profiles of proteins which were visualized by two-dimensional protein gels and autoradiography. The data indicated that eIF(iso)4E was particularly required for translation of the stored mRN...
Coordination of cell growth and cell division is very important for living organisms in order for... more Coordination of cell growth and cell division is very important for living organisms in order for these to develop harmonically. The present research is concerned with the purification and characterization of a new peptide hormone, namely ZmIGF (Zea mays insulin-like growth factor), which regulates growth and cell division in maize tissues. ZmIGF is a peptide of 5.7 kDa, as determined by mass spectroscopy. It was isolated either from maize embryonic axes of 48-h germinated seeds or from embryogenic callus and purified through several chromatographic procedures to obtain a single peak as shown by Reverse Phase High-Performance Liquid Chromatography (RP-HPLC). This peptide exhibits a well defined α-helix structure by circular dichroism analysis, similar to that reported for Insulin or for Insulin-like growth factor (IGF-1). Further, ZmIGF seems to perform, in maize, a similar function to that reported for insulin or peptides from the IGF family in animals. Indeed, maize tissues stimulated either by ZmIGF or insulin showed to induce selective synthesis of ribosomal proteins as well as of DNA. Taken together, the previously mentioned data strongly suggest that plants contain a peptide hormone of the IGF family, highly conserved through evolution that regulates growth and development.
ABSTRACT Pérez, L., Aguilar, R. and Sánchez-de-Jiménez, E. 1987. Effect of an exogenous auxin on ... more ABSTRACT Pérez, L., Aguilar, R. and Sánchez-de-Jiménez, E. 1987. Effect of an exogenous auxin on maize tissues. Alteration of protein synthesis and phosphorylation. - Physiol. Plantarum 69: 517–522. A synthetic auxin 2-(2-methyl-4-chloro)phenoxypropionic acid (MCPP), analogue of 2,4-D, alters maize (Zea mays L. H-30) germination while inducing callus formation. The effect of this auxin on protein synthesis and phosphorylation of the embryonic tissues was explored. Total cytoplasmic proteins were analysed for 14C or 32P incorporation into trichloroacetic acid precipitable material. MCPP significantly stimulated protein synthesis as well as protein phosphorylation. The protein synthesis pattern was highly altered in the presence of MCPP as analysed by two-dimensional gel electrophoresis. Analyses by Sephadex G-100 chromatography and by two-dimensional gel electrophoresis of phosphorylated proteins indicate that the effect of MCPP on protein phosphorylation was only quantitative.
Previous work in our lab suggests translational control during maize germination. To test this po... more Previous work in our lab suggests translational control during maize germination. To test this possibility the present research focuses on the phosphorylated status of the ribosomal proteins of maize axes during germination. Ribosomes from embryonic axes incubated for different periods were in vitro or in vivo labeled by 1h-pulse for ³²P orthophosphate. Electrophoretic analysis of the ribosomal proteins and autoradiographs revealed: (a) in vitro, several ³²P bands and very similar patterns for all stages tested (0 to 24h); in vivo, less number of labeled bands and a changing pattern from 3 to 24h of incubation. A protein of 30,900 MW did not appear phosphorylated until 8h of incubation, while a 17,000 MW protein was strongly labeled at 3h and fastly dephosphorylated toward 24h. Phosphorylated proteins belong to both the small and the large subunits. The implication of this process will be discussed.
In Vitro Cellular & Developmental Biology - Plant, 2010
Mechanisms that bring about coordination of cell growth and cell division in different organisms ... more Mechanisms that bring about coordination of cell growth and cell division in different organisms are biological events not yet clearly revealed. In maize, insulin effector of the phosphatidylinositol 3-kinase (PI3K)–target of rapamycin (TOR) signal transduction pathway in metazoan or an intrinsic maize growth factor similar to insulin has shown to regulate cell growth. This research has been undertaken to analyze
This article focuses on the effect that S6 ribosomal protein phosphorylation might have in regula... more This article focuses on the effect that S6 ribosomal protein phosphorylation might have in regulating mRNA translation. Maize axes of either 4 or 14 h of germination were pulse-labelled for 1 h with [32P]-orthophosphate. Analysis of their ribosomal proteins by gel electrophoresis and autoradiography showed distinctive levels of S6 ribosomal protein phosphorylation for both ribosomal sets. Axes at these two stages of germination were treated with alpha-amanitin to ensure transcription inhibition and pulse-labelling with [35S]-methionine. The [35S]-proteins, resulting from stored mRNA translation, when analysed by 2-D-gel electrophoresis and fluorography revealed distinctive [35S]-protein patterns. In vitro translation of stored mRNA on ribosomes from either 4 or 14 h germinated-maize axes produced different [35S]-protein patterns. Further, addition of 7methyl-GTP-Sepharose to the translation system showed differential cap-dependent protein synthesis inhibition depending on the set of ribosomes tested. It is concluded that translation of stored mRNA in germinating maize axes is at least partially regulated by a mechanism that involves S6 ribosomal protein phosphorylation.
Acidic ribosomal proteins (ARPs) are highly conserved phosphoproteins in eukaryotic organisms. Th... more Acidic ribosomal proteins (ARPs) are highly conserved phosphoproteins in eukaryotic organisms. They participate in translation regulation by interacting with eEF-2 elongation factors in the peptide elongation process. During maize germination, protein synthesis is tightly regulated by different mechanisms that are not yet clearly understood. The objective of this research is to characterize the expression patterns of the two maize ARPs (P1 and P2) and their phosphorylated status in germinating maize embryonic axes. Expression of P1 and P2 mRNA transcripts was analyzed by Northern blots with specific cDNA probes. Results indicated that both transcripts are among the mRNA stored pool of the quiescent axes and each displays a distinctive expression pattern during germination. P1 and P2 synthesis initiates very early in germination, as demonstrated by [(35)S]methionine pulse-labeling experiments. This synthesis was not insulin/IGF-stimulated as the synthesis of the bulk of ribosomal proteins that was responsive to this stimulus. P1 and P2 proteins were purified from ribosomes of maize embryonic axes and their physicochemical characteristics determined. A cytoplasmic pool of dephosphorylated P1 and P2 proteins was found in axes of quiescent and germinated stages that freely assembled into the ribosomes. IEF analysis of ARPs revealed one P1 (P1-1) and two P2 (P2-1 and P2-2) forms in the ribosomes of 24 h germinated axes. Kinetic studies of ARP phosphorylation during germination revealed a specific order of phospho-ARP appearance, suggesting that this process is under regulation within this period. It is concluded that P1 and P2 phosphorylation rather than ARP expression or assembly into ribosomes is the main step that regulates ARP function in axes during maize germination.
The primordial TOR pathway, known to control growth and cell proliferation, has still not been fu... more The primordial TOR pathway, known to control growth and cell proliferation, has still not been fully described for plants. Nevertheless, in maize, an insulin-like growth factor (ZmIGF) peptide has been reported to stimulate this pathway. This research provides further insight into the TOR pathway in maize, using a biochemical approach in cultures of fast-growing (FG) and slow-growing (SG) calli, as a model system. Our results revealed that addition of either ZmIGF or insulin to SG calli stimulated DNA synthesis and increased the growth rate through cell proliferation and increased the rate of ribosomal protein (RP) synthesis by the selective mobilization of RP mRNAs into polysomes. Furthermore, analysis of the phosphorylation status of the main TOR and S6K kinases from the TOR pathway revealed stimulation by ZmIGF or insulin, whereas rapamycin inhibited its activation. Remarkably, a putative maize insulin-like receptor was recognized by a human insulin receptor antibody, as demonstrated by immunoprecipitation from membrane protein extracts of maize callus. Furthermore, competition experiments between ZmIGF and insulin for the receptor site on maize protoplasts suggested structural recognition of the putative receptor by either effector. These data were confirmed by confocal immunolocalization within the cell membrane of callus cells. Taken together, these data indicate that cell growth and cell proliferation in maize depend on the activation of the TOR-S6K pathway through the interaction of an insulin-like growth factor and its receptor. This evidence suggests that higher plants as well as metazoans have conserved this biochemical pathway to regulate their growth, supporting the conclusion that it is a highly evolved conserved pathway.
... DOI: 10.1080/09291010400028542 Rebeca Toledo a c , Raúl Aguilar-Roblero b , Enrique Canchola ... more ... DOI: 10.1080/09291010400028542 Rebeca Toledo a c , Raúl Aguilar-Roblero b , Enrique Canchola c & Mario Caba Dr a * pages 47-55. Available online: 02 Sep 2011. ...
A characteristic mechanism of gene expression regulation during seed germination is the selective... more A characteristic mechanism of gene expression regulation during seed germination is the selective translation of mRNAs. Previous findings indicate that the two cap-binding complexes eIF4F (with eIF4E and eIF4G subunits) and eIF(iso)4F [with eIF(iso)4E and eIF(iso)4G subunits] are differentially expressed during maize seed germination. In addition, several studies in vitro have suggested that these factors may participate in selective mRNA translation. The translational activities of eIF4E and eIF(iso)4E were tested in vitro using transcripts from two different sets: dry (0 h) and 24-h-imbibed maize embryonic axes. In vitro translation of these mRNA pools in the presence of the recombinant eIF4E or eIF(iso)4E, and the native cap-binding complexes from dry- or 24-h-imbibed axes, produced different profiles of proteins which were visualized by two-dimensional protein gels and autoradiography. The data indicated that eIF(iso)4E was particularly required for translation of the stored mRN...
Coordination of cell growth and cell division is very important for living organisms in order for... more Coordination of cell growth and cell division is very important for living organisms in order for these to develop harmonically. The present research is concerned with the purification and characterization of a new peptide hormone, namely ZmIGF (Zea mays insulin-like growth factor), which regulates growth and cell division in maize tissues. ZmIGF is a peptide of 5.7 kDa, as determined by mass spectroscopy. It was isolated either from maize embryonic axes of 48-h germinated seeds or from embryogenic callus and purified through several chromatographic procedures to obtain a single peak as shown by Reverse Phase High-Performance Liquid Chromatography (RP-HPLC). This peptide exhibits a well defined α-helix structure by circular dichroism analysis, similar to that reported for Insulin or for Insulin-like growth factor (IGF-1). Further, ZmIGF seems to perform, in maize, a similar function to that reported for insulin or peptides from the IGF family in animals. Indeed, maize tissues stimulated either by ZmIGF or insulin showed to induce selective synthesis of ribosomal proteins as well as of DNA. Taken together, the previously mentioned data strongly suggest that plants contain a peptide hormone of the IGF family, highly conserved through evolution that regulates growth and development.
ABSTRACT Pérez, L., Aguilar, R. and Sánchez-de-Jiménez, E. 1987. Effect of an exogenous auxin on ... more ABSTRACT Pérez, L., Aguilar, R. and Sánchez-de-Jiménez, E. 1987. Effect of an exogenous auxin on maize tissues. Alteration of protein synthesis and phosphorylation. - Physiol. Plantarum 69: 517–522. A synthetic auxin 2-(2-methyl-4-chloro)phenoxypropionic acid (MCPP), analogue of 2,4-D, alters maize (Zea mays L. H-30) germination while inducing callus formation. The effect of this auxin on protein synthesis and phosphorylation of the embryonic tissues was explored. Total cytoplasmic proteins were analysed for 14C or 32P incorporation into trichloroacetic acid precipitable material. MCPP significantly stimulated protein synthesis as well as protein phosphorylation. The protein synthesis pattern was highly altered in the presence of MCPP as analysed by two-dimensional gel electrophoresis. Analyses by Sephadex G-100 chromatography and by two-dimensional gel electrophoresis of phosphorylated proteins indicate that the effect of MCPP on protein phosphorylation was only quantitative.
Previous work in our lab suggests translational control during maize germination. To test this po... more Previous work in our lab suggests translational control during maize germination. To test this possibility the present research focuses on the phosphorylated status of the ribosomal proteins of maize axes during germination. Ribosomes from embryonic axes incubated for different periods were in vitro or in vivo labeled by 1h-pulse for ³²P orthophosphate. Electrophoretic analysis of the ribosomal proteins and autoradiographs revealed: (a) in vitro, several ³²P bands and very similar patterns for all stages tested (0 to 24h); in vivo, less number of labeled bands and a changing pattern from 3 to 24h of incubation. A protein of 30,900 MW did not appear phosphorylated until 8h of incubation, while a 17,000 MW protein was strongly labeled at 3h and fastly dephosphorylated toward 24h. Phosphorylated proteins belong to both the small and the large subunits. The implication of this process will be discussed.
In Vitro Cellular & Developmental Biology - Plant, 2010
Mechanisms that bring about coordination of cell growth and cell division in different organisms ... more Mechanisms that bring about coordination of cell growth and cell division in different organisms are biological events not yet clearly revealed. In maize, insulin effector of the phosphatidylinositol 3-kinase (PI3K)–target of rapamycin (TOR) signal transduction pathway in metazoan or an intrinsic maize growth factor similar to insulin has shown to regulate cell growth. This research has been undertaken to analyze
This article focuses on the effect that S6 ribosomal protein phosphorylation might have in regula... more This article focuses on the effect that S6 ribosomal protein phosphorylation might have in regulating mRNA translation. Maize axes of either 4 or 14 h of germination were pulse-labelled for 1 h with [32P]-orthophosphate. Analysis of their ribosomal proteins by gel electrophoresis and autoradiography showed distinctive levels of S6 ribosomal protein phosphorylation for both ribosomal sets. Axes at these two stages of germination were treated with alpha-amanitin to ensure transcription inhibition and pulse-labelling with [35S]-methionine. The [35S]-proteins, resulting from stored mRNA translation, when analysed by 2-D-gel electrophoresis and fluorography revealed distinctive [35S]-protein patterns. In vitro translation of stored mRNA on ribosomes from either 4 or 14 h germinated-maize axes produced different [35S]-protein patterns. Further, addition of 7methyl-GTP-Sepharose to the translation system showed differential cap-dependent protein synthesis inhibition depending on the set of ribosomes tested. It is concluded that translation of stored mRNA in germinating maize axes is at least partially regulated by a mechanism that involves S6 ribosomal protein phosphorylation.
Acidic ribosomal proteins (ARPs) are highly conserved phosphoproteins in eukaryotic organisms. Th... more Acidic ribosomal proteins (ARPs) are highly conserved phosphoproteins in eukaryotic organisms. They participate in translation regulation by interacting with eEF-2 elongation factors in the peptide elongation process. During maize germination, protein synthesis is tightly regulated by different mechanisms that are not yet clearly understood. The objective of this research is to characterize the expression patterns of the two maize ARPs (P1 and P2) and their phosphorylated status in germinating maize embryonic axes. Expression of P1 and P2 mRNA transcripts was analyzed by Northern blots with specific cDNA probes. Results indicated that both transcripts are among the mRNA stored pool of the quiescent axes and each displays a distinctive expression pattern during germination. P1 and P2 synthesis initiates very early in germination, as demonstrated by [(35)S]methionine pulse-labeling experiments. This synthesis was not insulin/IGF-stimulated as the synthesis of the bulk of ribosomal proteins that was responsive to this stimulus. P1 and P2 proteins were purified from ribosomes of maize embryonic axes and their physicochemical characteristics determined. A cytoplasmic pool of dephosphorylated P1 and P2 proteins was found in axes of quiescent and germinated stages that freely assembled into the ribosomes. IEF analysis of ARPs revealed one P1 (P1-1) and two P2 (P2-1 and P2-2) forms in the ribosomes of 24 h germinated axes. Kinetic studies of ARP phosphorylation during germination revealed a specific order of phospho-ARP appearance, suggesting that this process is under regulation within this period. It is concluded that P1 and P2 phosphorylation rather than ARP expression or assembly into ribosomes is the main step that regulates ARP function in axes during maize germination.
The primordial TOR pathway, known to control growth and cell proliferation, has still not been fu... more The primordial TOR pathway, known to control growth and cell proliferation, has still not been fully described for plants. Nevertheless, in maize, an insulin-like growth factor (ZmIGF) peptide has been reported to stimulate this pathway. This research provides further insight into the TOR pathway in maize, using a biochemical approach in cultures of fast-growing (FG) and slow-growing (SG) calli, as a model system. Our results revealed that addition of either ZmIGF or insulin to SG calli stimulated DNA synthesis and increased the growth rate through cell proliferation and increased the rate of ribosomal protein (RP) synthesis by the selective mobilization of RP mRNAs into polysomes. Furthermore, analysis of the phosphorylation status of the main TOR and S6K kinases from the TOR pathway revealed stimulation by ZmIGF or insulin, whereas rapamycin inhibited its activation. Remarkably, a putative maize insulin-like receptor was recognized by a human insulin receptor antibody, as demonstrated by immunoprecipitation from membrane protein extracts of maize callus. Furthermore, competition experiments between ZmIGF and insulin for the receptor site on maize protoplasts suggested structural recognition of the putative receptor by either effector. These data were confirmed by confocal immunolocalization within the cell membrane of callus cells. Taken together, these data indicate that cell growth and cell proliferation in maize depend on the activation of the TOR-S6K pathway through the interaction of an insulin-like growth factor and its receptor. This evidence suggests that higher plants as well as metazoans have conserved this biochemical pathway to regulate their growth, supporting the conclusion that it is a highly evolved conserved pathway.
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