Estrés traumático secundario (ETS) en periodistas Mexicanos y defensores de derechos humanos / Secondary traumatic stress (STS) in Mexican journalists and human dights defenders

El objetivo del presente estudio descriptivo fue identificar la prevalencia de síntomas de estrés traumático secundario (ETS) en una muestra conjunta de periodistas mexicanos y defensores de derechos humanos (N igual 88), cuyo trabajo profesional demanda regularmente un contacto cercano con víctimas de violencia. Se encontró que 36.4 por ciento de los participantes presentaron sintomatología "alta” o “severa” de ETS. Sin embargo, no se ubicaron diferencias significativas entre ambos grupos. Por otra parte, las mujeres y quienes laboraban más de 40 horas a la semana, sí mostraron síntomas significativamente más altos. Los resultados de esta investigación transversal reflejan el considerable desgaste psicológico que pueden generar las exposiciones secundarias en profesionistas que documentan y establecen vínculos sistemáticos con personas traumatizadas por la violencia social en México.

The main goal of this descriptive study was to identify the prevalence of secondary traumatic stress (STS) symptoms in a pooled sample of Mexican journalists and human right defenders (N same 88), whose activities regularly demand a close contact with victims of violence. It was found that 36.4 percen of the participants presented “high” or “severe” STS symptoms. However, no significant differences between these groups of professionals were observed. Conversely, women and those who worked more than 40 hours a week presented significantly more severe symptoms. The results of this transversal investigation reflect the psychologic wear that these secondary exposures can generate in professionals who establish systematic links with subjects who have been traumatized by the social violence prevalent in modern Mexican society.

Percepción de la psiquiatría en estudiantes de pregrado de una facultad de medicina / Perception of Psychiatry among Undergraduate Students of a School of Medicine

Introducción: Los objetivos de este artículo son conocer las percepciones y creencias acerca de la psiquiatría en estudiantes de pregrado de medicina y comparar concepciones y apreciaciones sobre aspectos positivos y negativos antes y después de la formación específica en psiquiatría. Métodos: Estudio observacional, transversal; investigación cualitativa y cuantitativa, aplicando una encuesta a 90 estudiantes de pregrado de medicina de la Universidad de La sabana, en dos grupos: uno antes y otro después de recibir el curso de psiquiatría. Resultados: El 52,2% correspondió a semestres previos al curso de Psiquiatría; 25,5% mostró intención en especializarse en Psiquiatría antes del curso; este porcentaje bajó al 13,4% luego del curso. Se asocia la intención de no especializarse en psiquiatría con haber tenido el curso en esta área (prueba exacta de Fisher, p=0,042). Hay menos variabilidad del concepto de psiquiatría después del curso. La mayoría no se especializará en psiquiatría por interés en otras áreas. Antes del curso, los estudiantes resaltaron los aspectos biológicos de la enfermedad mental; después, prestan atención adicionalmente a otros factores. Antes y después del curso se considera que el manejo de estos pacientes es principalmente farmacológico. Se resalta el carácter incurable de las enfermedades mentales, el riesgo de enfermarse al atender los pacientes y la estigmatización. Conclusiones: La psiquiatría es concebida como una especialidad con énfasis en el tratamiento farmacológico. Hay baja frecuencia de estudiantes interesados en especializarse en esta área. El curso de psiquiatría se asocia con reducción de esta frecuencia y limita la variabilidad del concepto.

Objective: Learn about perceptions and beliefs regarding psychiatry among undergraduate medicine students and compare their conceptions and appreciations concerning positive and negative aspects, before and after specific training in psychiatry. Methods: Observational, cross-sectional study; qualitative and quantitative research with application of a survey for undergraduate medicine students of the Universidad de La Sabana, before and after a specific psychiatry course. 90 students answered the survey in two groups: one of them before the course, and the other one afterwards; Results: 52,2% corresponded to semesters prior to the course of psychiatry; 25.5% expressed the purpose to specialize in Psychiatry before the course, and such percentage decreased to 13.4% after the course. Association was found between the purpose of not specializing in Psychiatry with the fact of having taken said course (Fisher's exact test, p=0,042). Most students would not specialize in psychiatry because they are interested in other areas. Before the course, students made emphasis on the biological aspects of mental disease. After the course, they also directed their attention to other factors. The two groups believe that the management of these patients is mainly pharmacological. The incurable character of mental illness was also highlighted together with the risk of getting ill and the stigma it entails. Conclusions: Psychiatry is perceived as a medical specialization with emphasis on pharmacological treatment. There is a low frequency of students interested in this area. The course of psychiatry is associated with reduction of this frequency and limits the variability of the psychiatric concept.

[Perception of Psychiatry among Undergraduate Students of a School of Medicine]. / Percepción de la psiquiatría en estudiantes de pregrado de una facultad de medicina.

OBJECTIVE: Learn about perceptions and beliefs regarding psychiatry among undergraduate medicine students and compare their conceptions and appreciations concerning positive and negative aspects, before and after specific training in psychiatry. METHODS: Observational, cross-sectional study; qualitative and quantitative research with application of a survey for undergraduate medicine students of the Universidad de La Sabana, before and after a specific psychiatry course. 90 students answered the survey in two groups: one of them before the course, and the other one afterwards; RESULTS: 52,2% corresponded to semesters prior to the course of psychiatry; 25.5% expressed the purpose to specialize in Psychiatry before the course, and such percentage decreased to 13.4% after the course. Association was found between the purpose of not specializing in Psychiatry with the fact of having taken said course (Fisher's exact test, p=0,042). Most students would not specialize in psychiatry because they are interested in other areas. Before the course, students made emphasis on the biological aspects of mental disease. After the course, they also directed their attention to other factors. The two groups believe that the management of these patients is mainly pharmacological. The incurable character of mental illness was also highlighted together with the risk of getting ill and the stigma it entails. CONCLUSIONS: Psychiatry is perceived as a medical specialization with emphasis on pharmacological treatment. There is a low frequency of students interested in this area. The course of psychiatry is associated with reduction of this frequency and limits the variability of the psychiatric concept.

Autophosphorylation of carboxy-terminal residues inhibits the activity of protein kinase CK1alpha.

CK1 constitutes a protein kinase subfamily that is involved in many important physiological processes. However, there is limited knowledge about mechanisms that regulate their activity. Isoforms CK1delta and CK1epsilon were previously shown to autophosphorylate carboxy-terminal sites, a process which effectively inhibits their catalytic activity. Mass spectrometry of CK1alpha and splice variant CK1alphaL has identified the autophosphorylation of the last four carboxyl-end serines and threonines and also for CK1alphaS, the same four residues plus threonine-327 and serine-332 of the S insert. Autophosphorylation occurs while the recombinant proteins are expressed in Escherichia coli. Mutation of four carboxy-terminal phosphorylation sites of CK1alpha to alanine demonstrates that these residues are the principal but not unique sites of autophosphorylation. Treatment of autophosphorylated CK1alpha and CK1alphaS with lambda phosphatase causes an activation of 80-100% and 300%, respectively. Similar treatment fails to stimulate the CK1alpha mutants lacking autophosphorylation sites. Incubation of dephosphorylated enzymes with ATP to allow renewed autophosphorylation causes significant inhibition of CK1alpha and CK1alphaS. The substrate for these studies was a synthetic canonical peptide for CK1 (RRKDLHDDEEDEAMS*ITA). The stimulation of activity seen upon dephosphorylation of CK1alpha and CK1alphaS was also observed using the known CK1 protein substrates DARPP-32, beta-catenin, and CK2beta, which have different CK1 recognition sequences. Autophosphorylation effects on CK1alpha activity are not due to changes in Km(app) for ATP or for peptide substrate but rather to the catalytic efficiency per pmol of enzyme. This work demonstrates that CK1alpha and its splice variants can be regulated by their autophosphorylation status.

CK2alpha/CK1alpha chimeras are sensitive to regulation by the CK2beta subunit.

The effect of CK2beta on the activity of CK2alpha and other protein kinases that can bind this regulatory subunit is not fully understood. In an attempt to improve our understanding of this effect, chimeras of CK2alpha and CK1alpha have been constructed. These chimeras contain different portions of the CK2alpha amino terminal region that are involved in the interaction with CK2beta to form CK2 tetramers. In the case of chimeras 1 and 2, the portions of CK2alpha replace the corresponding segments of CK1alpha. In the case of chimera 3, the fragment of CK2alpha is added to the whole CK1alpha molecule with the exception of the initial methionine. Chimera 3 has 8% of the activity of CK1alphaWT, while chimeras 1 and 2 are 3 orders of magnitude less active than CK1alphaWT. All three chimeras bind tightly to CK2beta, but only chimeras 1 and 2 are significantly stimulated in their capacity to phosphorylate casein and canonical peptide substrates by addition of the regulatory subunit. No stimulation was observed with phosvitin or non-canonical peptides derived from beta-catenin. CK2beta protects chimeras 1 and 2 from thermal inactivation. Chimera 2 can phosphorylate CK2beta and autophosphorylate; however, salt concentrations above 150 mM NaCl eliminate the phosphorylation of CK2beta but not the autophosphorylation of chimera 2. Similarly, high salt decrease the stimulatory effect of CK2beta on the phosphorylation of casein.

Protein kinase CK2 as an ectokinase: the role of the regulatory CK2beta subunit.

Protein kinase CK2 (also known as casein kinase 2) is present in the cytoplasm, nuclei, and several other organelles. In addition, this enzyme has been found bound to the external side of the cell membrane where it acts as an ectokinase phosphorylating several extracellular proteins. Previous experiments with transfection of HEK-293T cells demonstrated that expression of both subunits, CK2alpha (catalytic) and CK2beta (regulatory), was necessary for the appearance of the ectopic enzyme as an ectokinase. In this work, using deletion and point mutations of CK2beta, it was possible to demonstrate that the region between amino acids 20 and 33 was necessary for the export of the enzyme as an ectokinase. Phenylalanines 21 and 22 and acidic residues in positions 26-28 are involved in the structural aspects that are required for export. However, the region encompassing amino acids 20-33 of CK2beta is not sufficient to make the carboxyl half of this subunit functional in bringing CK2 to the ectokinase locus. In cells transfected with only CK2beta, it was demonstrated that 3-4% of the subunit is exported to the cell medium, but the subunit is not bound to the external membrane.

Chemical dissection of the APC Repeat 3 multistep phosphorylation by the concerted action of protein kinases CK1 and GSK3.

A crucial event in machinery controlled by Wnt signaling is the association of beta-catenin with the adenomatous polyposis coli (APC) protein, which is essential for the degradation of beta-catenin and requires the multiple phosphorylation of APC at six serines (1501, 1503, 1504, 1505, 1507, and 1510) within its repeat three (R3) region. Such a phosphorylation is believed to occur by the concerted action of two protein kinases, CK1 and GSK3, but its mechanistic aspects are a matter of conjecture. Here, by combining the usage of variably phosphorylated peptides reproducing the APC R3 region and Edman degradation assisted localization of residues phosphorylated by individual kinases, we show that the process is initiated by CK1, able to phosphorylate S1510 and S1505, both specified by non-canonical determinants. Phosphorylation of S1505 primes subsequent phosphorylation of S1501 by GSK3. In turn, phospho-S1501 triggers the hierarchical phosphorylation of S1504 and S1507 by CK1. Once phosphorylated, S1507 primes the phosphorylation of both S1510 and S1503 by CK1 and GSK3, respectively, thus completing all six phosphorylation steps. Our data also rule out the intervention of CK2 despite the presence of a potential CK2 phosphoacceptor site, S1510LDE, in the R3 repeat. S1510 is entirely unaffected by CK2, while it is readily phosphorylated even in the unprimed peptide by CK1delta but not by CK1gamma. This discloses a novel motif significantly different from non-canonical sequences phosphorylated by CK1 in other proteins, which appears to be specifically recognized by the delta isoform of CK1.

Impaired hyperphosphorylation of rotavirus NSP5 in cells depleted of casein kinase 1alpha is associated with the formation of viroplasms with altered morphology and a moderate decrease in virus replication.

The rotavirus (RV) non-structural protein 5, NSP5, is encoded by the smallest of the 11 genomic segments and localizes in 'viroplasms', cytoplasmic inclusion bodies in which viral RNA replication and packaging take place. NSP5 is essential for the replicative cycle of the virus because, in its absence, viroplasms are not formed and viral RNA replication and transcription do not occur. NSP5 is produced early in infection and undergoes a complex hyperphosphorylation process, leading to the formation of proteins differing in electrophoretic mobility. The role of hyperphosphorylation of NSP5 in the replicative cycle of rotavirus is unknown. Previous in vitro studies have suggested that the cellular kinase CK1alpha is responsible for the NSP5 hyperphosphorylation process. Here it is shown, by means of specific RNA interference, that in vivo, CK1alpha is the enzyme that initiates phosphorylation of NSP5. Lack of NSP5 hyperphosphorylation affected neither its interaction with the virus VP1 and NSP2 proteins normally found in viroplasms, nor the production of viral proteins. In contrast, the morphology of viroplasms was altered markedly in cells in which CK1alpha was depleted and a moderate decrease in the production of double-stranded RNA and infectious virus was observed. These data show that CK1alpha is the kinase that phosphorylates NSP5 in virus-infected cells and contribute to further understanding of the role of NSP5 in RV infection.

The first armadillo repeat is involved in the recognition and regulation of beta-catenin phosphorylation by protein kinase CK1.

Multiple phosphorylation of beta-catenin by glycogen synthase kinase 3 (GSK3) in the Wnt pathway is primed by CK1 through phosphorylation of Ser-45, which lacks a typical CK1 canonical sequence. Synthetic peptides encompassing amino acids 38-64 of beta-catenin are phosphorylated by CK1 on Ser-45 with low affinity (K(m) approximately 1 mM), whereas intact beta-catenin is phosphorylated at Ser-45 with very high affinity (K(m) approximately 200 nM). Peptides extended to include a putative CK1 docking motif (FXXXF) at 70-74 positions or a F74AA mutation in full-length beta-catenin had no significant effect on CK1 phosphorylation efficiency. beta-Catenin C-terminal deletion mutants up to residue 181 maintained their high affinity, whereas removal of the 131-181 fragment, corresponding to the first armadillo repeat, was deleterious, resulting in a 50-fold increase in K(m) value. Implication of the first armadillo repeat in beta-catenin targeting by CK1 is supported in that the Y142E mutation, which mimics phosphorylation of Tyr-142 by tyrosine kinases and promotes dissociation of beta-catenin from alpha-catenin, further improves CK1 phosphorylation efficiency, lowering the K(m) value to <50 nM, approximating the physiological concentration of beta-catenin. In contrast, alpha-catenin, which interacts with the N-terminal region of beta-catenin, prevents Ser-45 phosphorylation of CK1 in a dose-dependent manner. Our data show that the integrity of the N-terminal region and the first armadillo repeat are necessary and sufficient for high-affinity phosphorylation by CK1 of Ser-45. They also suggest that beta-catenin association with alpha-catenin and beta-catenin phosphorylation by CK1 at Ser-45 are mutually exclusive.

Identifying interaction motifs in CK2beta--a ubiquitous kinase regulatory subunit.

Casein kinase 2 (CK2) is probably the most ubiquitous serine/threonine kinase found in eukaryotes: it phosphorylates >300 cellular proteins, ranging from transcription factors to proteins involved in chromatin structure and cell division. CK2 is a heterotetrameric enzyme that induces neoplastic growth when overexpressed. The beta subunit of CK2 (CK2beta) functions as the regulator of the catalytic CK2alpha and CK2alpha' subunits, enhancing their stability, activity and specificity. However, CK2beta also functions as a multisubstrate docking platform for several other binding partners. Here, we discuss the organization and roles of interaction motifs of CK2beta, postulate new protein-interaction sites and map these to the known interaction motifs, and show how the resulting complexity of interactions mediated by CK2 gives rise to the versatile functions of this pleiotropic protein kinase.

Casein kinase 2-dependent serine phosphorylation of MuSK regulates acetylcholine receptor aggregation at the neuromuscular junction.

The release of Agrin by motoneurons activates the muscle-specific receptor tyrosine kinase (MuSK) as the main organizer of subsynaptic specializations at the neuromuscular junction. MuSK downstream signaling is largely undefined. Here we show that protein kinase CK2 interacts and colocalizes with MuSK at post-synaptic specializations. We observed CK2-mediated phosphorylation of serine residues within the kinase insert (KI) of MuSK. Inhibition or knockdown of CK2, or exchange of phosphorylatable serines by alanines within the KI of MuSK, impaired acetylcholine receptor (AChR) clustering, whereas their substitution by residues that imitate constitutive phosphorylation led to aggregation of AChRs even in the presence of CK2 inhibitors. Impairment of AChR cluster formation after replacement of MuSK KI with KIs of other receptor tyrosine kinases correlates with potential CK2-dependent serine phosphorylation within KIs. MuSK activity was unchanged but AChR stability decreased in the presence of CK2 inhibitors. Muscle-specific CK2beta knockout mice develop a myasthenic phenotype due to impaired muscle endplate structure and function. This is the first description of a regulatory cross-talk between MuSK and CK2 and of a role for the KI of the receptor tyrosine kinase MuSK for the development of subsynaptic specializations.

Minimal CK2 activity required for yeast growth.

Protein kinase CK2 is essential for the growth of Saccharomyces cerevisiae. Yeast cells that lack the functional genes coding for both the catalytic subunits of protein kinase CK2 can grow only if they are complemented by exogenous cDNAs coding for this subunit. A series of deletion mutants of CK2alpha from Xenopus laevis was constructed. These mutants that have carboxyl end deletions yield a CK2alpha product that varies over four orders of magnitude in its capacity to phosphorylate casein in vitro. Complementation of yeast RPG41-1a, a mutant defective in CKA1 and CKA2 genes, with wild-type X. laevis CK2alpha and with cDNAs coding for truncated CK2alpha having amino acids 1-328 and 1-327 resulted in cells that grew in gal-minimal media at 30 degrees C as well as the cells harboring the yeast CKA2 gene. However, the growth was significantly diminished when cells were complemented with X. laevis CK2alpha containing 1-326 amido acids. This mutant has 0.6% of the catalytic activity of the wild-type enzyme. Yeast cells that expressed CK2alpha 1-324 and 1-323 which have 10-and 100-fold less activity, respectively, were not able to grow. The growth of cells containing the CK2alpha 1-326 mutant was very sensitive to temperature, and minimal growth was observed at 37 degrees C. This mutant was also more sensitive to UV radiation but was not significantly affected by 0.4 M NaCl.

Generation of protein kinase Ck1alpha mutants which discriminate between canonical and non-canonical substrates.

Protein kinase CK1 denotes a family of pleiotropic serine/threonine protein kinases implicated in a variety of cellular functions. Typically, CK1 acts as a 'phosphate-directed' kinase whose targeting is primed by a single phosphorylated side chain at position n-3 or n-4 relative to serine/threonine, but increasing evidence is accumulating that CK1 can also engage some of its substrates at sites that do not conform to this canonical consensus. In the present paper, we show that CK1a phosphorylates with the same efficiency phosphopeptides primed by a phosphoserine residue at either n-3 [pS(-3)] or n-4 [pS(-4)] positions. The phosphorylation efficiency of the pS(-4) peptide, and to a lesser extent that of the pS(-3) peptide, is impaired by the triple mutation of the lysine residues in the K229KQK232 stretch to alanine residues, promoting 40-fold and 6-fold increases of Km respectively. In both cases, the individual mutation of Lys232 is as detrimental as the triple mutation. A kinetic alanine-scan analysis with a series of substituted peptide substrates in which the priming phosphoserine residue was effectively replaced by a cluster of four aspartate residues was also consistent with a crucial role of Lys232 in the recognition of the acidic determinant at position n-4. In sharp contrast, the phosphorylation of b-catenin and of a peptide including the non-canonical b-catenin site (Ser45) lacking acidic/phosphorylated determinants upstream is not significantly affected by mutations in the KKQK stretch. These data provide a molecular insight into the structural features that underlie the site specificity of CK1a and disclose the possibility of developing strategies for the preferential targeting of subsets of CK1 substrates.

Protein kinase casein kinase 2 holoenzyme produced ectopically in human cells can be exported to the external side of the cellular membrane.

Ectokinases can phosphorylate extracellular proteins and external domains of membrane proteins influencing cell adhesion, movement, and cellular interactions. An ectokinase with the properties of casein kinase 2 (CK2) has been previously described, but little is known about the structural characteristics that allow this enzyme to be exported from the cell. Transfection of human embryonic kidney-293 cells with cDNAs coding for the catalytic (CK2alpha or CK2alpha') and regulatory (CK2beta) subunits with hemaglutinin tags allowed us to study the export of ectopically synthesized enzyme. When the catalytic (CK2alpha or CK2alpha') and the CK2beta regulatory subunits are cotransfected, the tetrameric enzyme composed of both subunits (holoenzyme) is detected outside the cell. This observation has been confirmed by assaying protein kinase activity in immunoprecipitates obtained with antihemaglutinin antibody by using a CK2-specific peptide substrate and by Western blots as well as by immunofluorescence of nonpermeabilized cells. Transfection with cDNA of catalytic or regulatory subunit alone does not result in export of these subunits. A study of the kinetics of appearance of the ectopically synthesized protein at different times after transfection indicates that a 5- to 7-h delay after the synthesis of the protein before it appears in the extracellular compartment. Using mutations of CK2alpha that eliminate phosphorylating activity [CK2alpha(Asp-156-Ala)] or that make it less sensitive to heparin inhibition [CK2alpha(Lys-75-Glu,Lys-76-Glu)] demonstrated that these mutations do not prevent the holoenzyme to be exported from the cells.

Basic region of residues 228-231 of protein kinase CK1alpha is involved in its interaction with axin: binding to axin does not affect the kinase activity.

Protein kinase CK1, also known as casein kinase 1, participates in the phosphorylation of beta-catenin, which regulates the functioning of the Wnt signaling cascade involved in embryogenesis and carcinogenesis. beta-catenin phosphorylation occurs in a multiprotein complex assembled on the scaffold protein axin. The interaction of CK1alpha from Danio rerio with mouse-axin has been studied using a pull-down assay that uses fragments of axin fused to glutathione S transferase, which is bound to glutathione sepharose beads. The results indicate that the three lysines present in the basic region of residues 228-231 of CK1alpha are necessary for the binding of CK1 to axin. Lysine 231 is particularly important in this interaction. In order to define the relevance of the axin-CK1alpha interaction, the effect of the presence of axin on the phosphorylating activity of CK1alpha was tested. It is also evident that the region of axin downstream of residues 503-562 is required for CK1alpha interaction. The binding of CK1alpha to axin fragment 292-681 does not facilitate the phosphorylation of beta-catenin despite the fact that this axin fragment can also bind beta-catenin. Binding of CK1alpha to axin is not required for the phosphorylation of axin itself and, likewise, axin does not affect the kinetic parameters of the CK1alpha towards casein or a specific peptide substrate.

Uncoupling substrate and activation functions of rotavirus NSP5: phosphorylation of Ser-67 by casein kinase 1 is essential for hyperphosphorylation.

Rotavirus NSP5 is a nonstructural protein that localizes in viroplasms of virus-infected cells. NSP5 interacts with NSP2 and undergoes a complex posttranslational hyperphosphorylation, generating species with reduced PAGE mobility. Here we show that NSP5 operates as an autoregulator of its own phosphorylation as a consequence of two distinct activities of the protein: substrate and activator. We developed an in vivo hyperphosphorylation assay in which two NSP5 mutant constructs are cotransfected. One of them, fused to an 11-aa tag, served as substrate whereas the other was used to map NSP5 domains required for activation. The activation and substrate activity could be uncoupled, demonstrating a hyperphosphorylation process in trans between the activator and substratum. This process involved dimerization of the two components through the 18-aa C-terminal tail. Phosphorylation of Ser-67 within the SDSAS motif (amino acids 63-67) was required to trigger hyperphosphorylation by promoting the activation function. We present evidence of casein kinase 1alpha being the protein kinase responsible for this key step as well as for the consecutive ones leading to NSP5 hyperphosphorylation.

Structure of the regulatory subunit of CK2 in the presence of a p21WAF1 peptide demonstrates flexibility of the acidic loop.

A truncated form of the regulatory subunit of the protein kinase CK2beta (residues 1-178) has been crystallized in the presence of a fragment of the cyclin-dependent kinase inhibitor p21WAF1 (residues 46-65) and the structure solved at 2.9 A resolution by molecular replacement. The core of the CK2beta dimer shows a high structural similarity with that identified in previous structural analyses of the dimer and the holoenzyme. However, the electron density corresponding to the substrate-binding acidic loop (residues 55-64) indicates two conformations that differ from that of the holoenzyme structure [Niefind et al. (2001), EMBO J. 20, 5320-5331]. Difference electron density near the dimerization region in each of the eight protomers in the asymmetric unit is attributed to between one and eight amino-acid residues of a complexed fragment of p21WAF1. This binding site corresponds to the solvent-accessible part of the conserved zinc-finger motif.

Cell cycle regulatory protein p27KIP1 is a substrate and interacts with the protein kinase CK2.

The protein kinase CK2 is constituted by two catalytic (alpha and/or alpha') and two regulatory (beta) subunits. CK2 phosphorylates more than 300 proteins with important functions in the cell cycle. This study has looked at the relation between CK2 and p27(KIP1), which is a regulator of the cell cycle and a known inhibitor of cyclin-dependent kinases (Cdk). We demonstrated that in vitro recombinant Xenopus laevis CK2 can phosphorylate recombinant human p27(KIP1), but this phosphorylation occurs only in the presence of the regulatory beta subunit. The principal site of phosphorylation is serine-83. Analysis using pull down and surface plasmon resonance (SPR) techniques showed that p27(KIP1) interacts with the beta subunit through two domains present in the amino and carboxyl ends, while CD spectra showed that p27(KIP1) phosphorylation by CK2 affects its secondary structure. Altogether, these results suggest that p27(KIP1) phosphorylation by CK2 probably involves a docking event mediated by the CK2beta subunit. The phosphorylation of p27(KIP1) by CK2 may affect its biological activity.

p53 codon 72 polymorphism and risk of cervical cancer.

Storey et al. (1998) implicated the proline/argine polymorphism of the codon 72 of the tumor-suppressor gene p53 in the development of cervical cancer (CC) with the observation that the p53 protein is more efficiently inactivated by the E6 oncoprotein of human papillomavirus in p53 arginine as compared with its proline isoform. These authors further noted that in the United Kingdom, individuals homozygous for the arginine allele were several times more susceptible to HPV-associated tumorigenesis that proline/arginine heterozygotes. Subsequent studies in different countries failed to unanimously confirm this association. Motivated by the high incidence of CC in Chile, we undertook a case control study obtaining the following frequencies for genotypes PP, AP and AA in 60 ICC cases and 53 carefully selected controls: 0.067, 0.250, 0.683 and 0.075, 0.453, 0.472 respectively. A significant difference (X2 = 3.19 p < 0.02) and an odds ratio of 2.62 supported Storey et al (1998)'s results. In addition, rejecting previous hypotheses about the world distribution of the p53 codon 72 polymorphism, we conclude that this distribution most likely represents ancient human dispersal routes. Several methodological and biological explanations for the results obtained in previous negative association studies are briefly discussed.