Iis - Integrated Interactome System: A Web-based Platform For The Annotation, Analysis And Visualization Of Protein-metabolite-gene-drug Interactions By Integrating A Variety Of Data Sources And Tools

Background: High-throughput screening of physical, genetic and chemical-genetic interactions brings important perspectives in the Systems Biology field, as the analysis of these interactions provides new insights into protein/gene function, cellular metabolic variations and the validation of therapeutic targets and drug design. However, such analysis depends on a pipeline connecting different tools that can automatically integrate data from diverse sources and result in a more comprehensive dataset that can be properly interpreted. Results: We describe here the Integrated Interactome System (IIS), an integrative platform with a web-based interface for the annotation, analysis and visualization of the interaction profiles of proteins/genes, metabolites and drugs of interest. IIS works in four connected modules: (i) Submission module, which receives raw data derived from Sanger sequencing (e.g. two-hybrid system); (ii) Search module, which enables the user to search for the processed reads to be assembled into contigs/singlets, or for lists of proteins/genes, metabolites and drugs of interest, and add them to the project; (iii) Annotation module, which assigns annotations from several databases for the contigs/singlets or lists of proteins/genes, generating tables with automatic annotation that can be manually curated; and (iv) Interactome module, which maps the contigs/singlets or the uploaded lists to entries in our integrated database, building networks that gather novel identified interactions, protein and metabolite expression/concentration levels, subcellular localization and computed topological metrics, GO biological processes and KEGG pathways enrichment. This module generates a XGMML file that can be imported into Cytoscape or be visualized directly on the web. Conclusions: We have developed IIS by the integration of diverse databases following the need of appropriate tools for a systematic analysis of physical, genetic and chemical-genetic interactions. IIS was validated with yeast two-hybrid, proteomics and metabolomics datasets, but it is also extendable to other datasets. 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Kinase Inhibitor Profile For Human Nek1, Nek6, And Nek7 And Analysis Of The Structural Basis For Inhibitor Specificity

Human Neks are a conserved protein kinase family related to cell cycle progression and cell division and are considered potential drug targets for the treatment of cancer and other pathologies. We screened the activation loop mutant kinases hNek1 and hNek2, wild-type hNek7, and five hNek6 variants in different activation/phosphorylation statesand compared them against 85 compounds using thermal shift denaturation. We identified three compounds with significant Tm shifts: JNK Inhibitor II for hNek1(Ä262-1258)-(T162A), Isogranulatimide for hNek6(S206A), and GSK-3 Inhibitor XIII for hNek7wt. Each one of these compounds was also validated by reducing the kinases activity by at least 25%. The binding sites for these compounds were identified by in silico docking at the ATP-binding site of the respective hNeks. Potential inhibitors were first screened by thermal shift assays, had their efficiency tested by a kinase assay, and were finally analyzed by molecular docking. 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Contribuição da heterogeneidade de linhas de regressão para a recomendação de novas cultivares

O objetivo deste trabalho foi evidenciar a contribuição da análise da heterogeneidade das linhas de regressão para a recomendação de novas cultivares, pela metodologia de Eberhart & Russell. Um experimento com dez genótipos de arroz, avaliados em oito ambientes, foi utilizado para ilustração da metodologia. Utilizou-se o delineamento de blocos ao acaso com três repetições. A soma dos quadrados da interação genótipo x ambiente (GxA) foi decomposta para avaliação da heterogeneidade das linhas de regressão e dos desvios acumulados da linearidade. A heterogeneidade das linhas de regressão foi analisada com o uso do teste t sobre os coeficientes de regressão linear dos genótipos. Os dois componentes ortogonais da interação GxA foram significativos. A análise da heterogeneidade das linhas de regressão permitiu detectar incoerências na adaptabilidade dos genótipos, o que diminui as chances de recomendações equivocadas de cultivares. Os genótipos foram classificados de acordo com a eficiência da metodologia de Eberhart & Russell em explicar a natureza do desempenho genotípico diante das mudanças nos ambientes. A avaliação da heterogeneidade das linhas de regressão contribui para a recomendação mais efetiva de novas cultivares com a metodologia de Eberhart & Russell

Treatment Of Varicose Veins With Endovenous Laser: A Prospective 39-month Follow-up Study [tratamento De Varizes Com Laser Endovenoso: Estudo Prospectivo Com Seguimento De 39 Meses]

Objective: To evaluate results of endovenous laser treatment for primary varicose veins. Patients and method: From July 2001 to September 2004 (39 months), 253 outpatients (417 limbs) were treated with 810 and 980 nm diode laser energy delivered percutaneously using optical fiber introduced by puncture under ultrasound guidance. Tumescent anesthesia (50-150 ml of 0.2% lidocaine) was delivered perivenously. Power and duration of the pulse were determined by vein diameter. Duplex control was performed at 7 days, 1 month, 3 months, 6 months, 1 year and yearly thereafter to assess treatment efficacy and adverse effects. Results: Primary great saphenous vein occlusion was obtained in 405 of 417 members (97.1%). Twelve recurrent cases (2.9%) were successfully treated. Mean follow-up time was 18 months. During this period, global recurrence rate of varicose veins was 7.4%; 6.3% (26 limbs) related to tributary and collateral veins of the saphenofemoral junction, and 1.2% (five limbs) with great saphenous vein recanalization. All recurrences occurred between 3 and 12 months. Ecchymosis was the most common adverse effect (60.6%). Other complications were: moderate pain during the procedure (16.1%); hematoma (5.5%), superficial phlebitis of varicose tributaries (3.4%), hyperpigmentation (2.9%), transient paresthesia (3.4%). There were no cases of great saphenous vein thrombophlebitis, deep vein thrombosis or pulmonary emboli. Conclusion: Varicose vein treatment with endovenous laser technique was successful in occluding great saphenous vein and its branches, with self-limited adverse effects and recurrence rate lower than 8% in the follow-up period. Copyright © 2006 by Sociedade Brasileira de Angiologia e de Cirurgia Vascular.53184193Maffei, F.H., Magaldi, C., Pinho, S.Z., Varicose veins and chronic venous insufficiency in Brazil: Prevalence among 1755 inhabitants of a country town (1986) Int J Epidemiol, 15, pp. 210-217Bergan, J.J., Ballard, J.I., Correction of superficial reflux (1998) Atlas of Endoscopic Perforator Vein Surgery, pp. 98-103. , Gloviczki P, Bergan JJ, editors. London: SpringerLofgren, E.P., Lofgren, K.A., Recurrence of varicose veins after the stripping operation (1971) Arch Surg, 102, pp. 111-114Dodd, H., Cockett, F.B., (1976) The Pathology and Surgery of the Veins of the Lewer Limb. 2nd Ed., pp. 210-214. , London: LivingstoneDavies, A.H., Steffen, C., Cosgrove, C., Wilkins, D.C., Varicose vein surgery: Patient satisfaction (1995) J R Coll Surg Edinb, 40, pp. 298-299Navarro, L., Min, R.J., Bone, C., Endovenous Laser: A new minimally invasive method of treatment of varicose veins-preliminary observations using an 810 nm diode laser (2001) Dermatol Surg, 27, pp. 117-122Proebstle, T.M., Lehr, H.A., Kargl, A., Endovenous treatment of the greater saphenous vein with a 940-nm diode laser: Thrombotic occlusion after endoluminal thermal damage by laser- generated steam bubbles (2002) J Vasc Surg, 35, pp. 729-736Timperman, P.E., Sichlau, M., Ryu, R.K., Greater energy delivery improves treatment success of endovenous laser treatment of incompetent saphenous veins (2004) J Vasc Interv Radiol, 15, pp. 1061-1063Dwerryhouse, S., Davies, B., Harradine, K., Earnshaw, J.J., Stripping the long saphenous vein reduces the rate of reoperation for recurrent varicose veins: Five-year results of a randomized trial (1999) J Vasc Surg, 29, pp. 589-592Manfrini, S., Gasbarro, V., Danielsson, G., Endovenous management of saphenous vein reflux (2000) J Vasc Surg, 32, pp. 330-342. , Endovenous Reflux Management Study GroupGlass, G.M., Neovascularisation in recurrence of the varicose great saphenous vein following transection (1987) Phlebology, 2, pp. 81-92Nyamekye, I., Shephard, N.A., Davies, B., Heather, B.P., Earnshaw, J.J., Clinicopathological evidence that neovascularisation is a cause of recurrent varicose veins (1998) Eur J Vasc Endovasc Surg, 15, pp. 412-415Van Rij, A.M., Jones, G.T., Hill, G.B., Jiang, P., Neovascularization and recurrent varicose veins: More histologic and ultrasound evidence (2004) J Vasc Surg, 40, pp. 296-302Fischer, R., Linde, N., Duff, C., Jeanneret, C., Chandler, J.G., Seeber, P., Late recurrent saphenofemoral junction reflux after ligation and stripping of the greater saphenous vein (2001) J Vasc Surg, 34, pp. 236-240Creton, D., Prosthetic material interposition on the crossectomy stump in varicose vein recurrence surgery: Preliminary report on the prevention of angiogenesis (1998) Scripta Phlebol, 6, pp. 4-7Glass, G.M., Prevention of saphenofemoral and saphenopopliteal recurrence of varicose veins by forming a partition to contain neovascularisation (1998) Phebology, 13, pp. 3-9Jakobsen, B.H., The value of different forms of treatment for varicose veins (1979) Br J Surg, 66, pp. 182-184Proebstle, T.M., Gul, D., Lehr, H.A., Kargl, A., Knop, J., Infrequent early recanalization of greater saphenous vein after endovenous laser treatment (2003) J Vas Surg, 38, pp. 511-516Min, R.J., Khilnani, N., Zimmet, S.E., Endovenous laser treatment of saphenous vein reflux: Long-term results (2003) J Vasc Interv Radiol, 14, pp. 991-996Timperman, P.E., Arteriovenous fistula after endovenous laser treatment of the short saphenous vein (2004) J Vasc Interv Radiol, 15, pp. 625-627Min, R.J., Zimmet, S.E., Isaacs, M.N., Forrestal, M.D., Endovenous laser treatment of the incompetent greater saphenous vein (2001) J Vasc Interv Radiol, 12, pp. 1167-1171Gerard, J.L., Desgranges, P., Becquemin, J.P., Desse, H., Melliere, D., Peut-on traiter les grandes saphénes variqueuses par laser endoveineaux en ambulatoire (2002) J Mal Vasc, 27, pp. 222-225Chang, C.J., Chua, J.J., Endovenous laser photocoagulation (EVLP) for varicose veins (2002) Lasers Surg Med, 31, pp. 257-262Oh, C.K., Jung, D.S., Jang, H.S., Kwon, K.S., Endovenous laser surgery of the incompetent greater saphenous vein with a 980 nm diode laser (2003) Dermatol Surg, 29, pp. 1135-1140Perkowski, P., Ravi, R., Gowda, R.C., Endovenous laser ablation of the saphenous vein for treatment of venous insufficiency and varicose veins: Early results from a large single-center experience (2004) J Endovasc Ther, 11, pp. 132-138Soracco, J.E., D'Ámbola, J.L., Ciucci, J.L., Godoy, J.M.P., Belczak, C.E.Q., Complicações no tratamento com laser endovascular em varizes de membros inferiores (2005) J Vasc Bras, 4, pp. 333-335Mozes, G., Kalra, M., Carmo, M., Swenson, L., Gloviczki, P., Extension of saphenous thrombus into the femoral vein: A potential complication of new endovenous ablation techniques (2005) J Vasc Surg, 41, pp. 130-135Goldman, M.P., Mauricio, M., Rao, J., Intravascular 1320-nm laser closure of the great saphenous vein: A 6- to 12-month follow-up study (2004) Dermatol Surg, 30, pp. 1380-1385Proebstle, T.M., Moehler, T., Gül, D., Herdemann, S., Endovenous treatment of the great saphenous vein using a 1,320 nm Nd:YAG laser causes fewer side effects than using a 940 nm diode laser (2005) Dermatol Surg, 31, pp. 1678-1683. , discussion 1683-4Agus, G.B., Mancini, S., Magi, G., The first 1000 cases of Italian Endovenous-Laser Working Group (IEWG). Rationale, and long-term outcomes for the 1999-2003 period (2006) Int Angiol, 25, pp. 209-215. , IEWGLisbona, C., Pérez, P., Segura, J., Puncernau, X., Laser surgery in venous insufficiency: E.L.A.V. personal technique (2004) J Vasc Bras, 3, pp. 403-406Keith Jr., L.M., Smead, W.L., Saphenous vein stripping and its complications (1983) Surg Clin North Am, 63, pp. 1303-1312Lofgren, E.P., Coates, H.L., O'Brien, P.C., Clinically suspect pulmonary embolism after vein stripping (1976) Mayo Clin Proc, 51, pp. 77-80Bounameaux, H., Huber, O., Postoperative deep vein thrombosis and surgery for varicose veins (1996) BMJ, 312, p. 115

Prevalence Of Abdominal Aortic Dilatation In Patients Aged 60 Years Or Older With Coronary Disease [prevalência De Dilatação Da Aorta Abdominal Em Coronariopatas Idosos]

Background: Indiscriminate screening programs for abdominal aortic aneurysm will help a small percentage of individuals. However, when considering groups with risk factors associated with aortic dilatation, which increases the probability of the disease, such programs will provide an adequate allocation of resources and a greater benefit to the population. Programs guided by medical societies, providing an early diagnosis of vascular diseases and consequently a better preparation of patients, would result in better survival rates with lower morbidity. Objective: To evaluate the prevalence of abdominal aortic dilatation in patients aged 60 years or older with atherosclerotic coronary disease diagnosed by coronary angiography. Methods: The sample selected for this study considered the fact that preoperative assessment of vascular surgery had not been indication for catheterization. Evaluation was then performed, based on anamnesis, physical examination and ultrasound Doppler of the abdominal aorta. Statistical analysis started with chi-square test, with further multivariate logistic regression analysis and univariate logistic regression, with p ≤ 0.05 considered significant. Results: Of 180 patients, 57 (31.7%) were female and 123 (68.3%) were male. Agevaried from 60 to 80 years, with mean of 66.7 years. Among the 16 individuals with abdominal aortic dilatation (10 aneurysms and six dilatations), only onewas female. The risk of an individual with one atherosclerotic coronary lesion presenting abdominal aortic dilatation was 0.4%. Similarly, in those with two or three lesions the risk was 1.7, and 4.5% in those with more than three lesions. When associated with smoking, these values were 6.9, 11.8 and 27.1%, respectively. Conclusion: The present study leads to the conclusion that prevalence of abdominal aortic dilatation was 8.9% (16 out of 180 patients) in this specific sample. It was more prevalent in males, smokers and associated with presence of diffuse atherosclerotic lesions of the coronary arteries. Copyright © 2007 by Sociedade Brasileira de Angiologia e de Cirurgia Vascular.62114123Matas R. Aneurismas. In: Martorell F. Angiologia: enfermedades vasculares. Barcelona: Salvat1972. p. 203-8Grande, R.F., Aspectos históicos de la cirugía de los aneurismas. Matas y las nuevas técnicas (1997) Patología Vasc, 3, pp. 75-87Kruppski WC. Arterial aneurysms. In: Rutherford RB. Vascular surgery. 4th ed. v. 2. Philadelphia: Saunders1995. p. 1025-32Brito JC. História da cirurgia dos aneurismas da aorta abdominal. In: Bonamigo TP, Von Ristow A. Aneurismas. Rio de Janeiro: Di livros2000. p. 23-30Gama DA. Passado, presente e futuro do tratamento do aneurisma da aorta abdominal. In: Bonamigo TP, Von Ristow A. Aneurismas. Rio de Janeiro: Di livros2000. p. 31-8Harris, L.M., Faggioli, G.L., Fiedler, R., Curl, G.R., Ricotta, J.J., Ruptured abdominal aortic aneurysms: Factors affecting mortality rates (1991) J Vasc Surg, 14, pp. 812-818. , discussion 819-20Gloviczki, P., Pairolero, P.C., Mucha Jr., P., Ruptured abdominal aortic aneurysms: Repair should not be denied (1992) J Vasc Surg, 15, pp. 851-857. , discussion 857-9Ernst, C.B., Abdominal aortic aneurysm (1993) N Engl J Med, 328, pp. 1167-1172Johansson, G., Swedenborg, J., Ruptured abdominal aortic aneurysms: A study of incidence and mortality (1986) Br J Surg, 73, pp. 101-103Collin, J., Araujo, L., Walton, J., Lindsell, D., Oxford screening programme for abdominal aortic aneurysm in men aged 65 to 74 years (1988) Lancet, 2, pp. 613-615Karkos, C.D., Mukhopadhyay, U., Papakostas, I., Ghosh, J., Thomson, G.J., Hughes, R., Abdominal aortic aneurysm: The role of clinical examination and opportunistic detection (2000) Eur J Vasc Endovasc Surg, 19, pp. 299-303Bonamigo TP. Tratamento de urgência nos aneurismas da aorta abdominal. In: Bonamigo TP, Burihan E, Cinelli Jr. M, von Ristow A. Doença da aorta e seus ramos. São Paulo: Fundo Editorial Byk1991. p. 48-58Petersen, M.J., Cambria, R.P., Kaufman, J.A., Magnetic resonance angiography in the preoperative evaluation of abdominal aortic aneurysms (1995) J Vasc Surg, 21, pp. 891-898. , discussion 899Van Bellen B. Diagnóstico por imagem do aneurisma da aorta abdominal. In: Bonamigo TP, Von Ristow A. Aneurismas. Rio de Janeiro: Dilivros2000. pRacy, D.J., Angiorressonância magnética de artérias periféricas (2002) Doenças vasculares periféricas, 1, pp. 441-453. , Maffei FHA, Lástória S, Yoshida WB, Rollo HA, Rio de Janeiro: Medsis;Faria, R.C.S., Tomografia computadorizada espiral nas patologias vasculares periféricas (2002) Doenças vasculares periféricas, 1, pp. 398-440. , Maffei FHA, Lástória S, Yoshida WB, Rollo HA, Rio de Janeiro: Medsis;Wassef, M., Baxter, B.T., Chisholm, R.L., Pathogenesis of abdominal aortic aneurysms: A multidisciplinary research program supported by the National Heart, Lung and Blood Institute (2001) J Vasc Surg, 34, pp. 730-738Hollier, L.H., Plate, G., O'Brien, P.C., Late survival after abdominal aortic aneurysm repair: Influence of coronary artery disease (1984) J Vasc Surg, 1, pp. 290-299Hertzer, N.R., Clinical experience with preoperative coronary angiography (1985) J Vasc Surg, 2, pp. 510-514Tilson, D., Status of research on abdominal aortic aneurysm disease (1989) J Vasc Surg, 9, pp. 367-369Johnston, K.W., Multicenter prospective study of nonruptured abdominal aortic aneurysm. Part II. Variables predicting morbidity and mortality (1989) J Vasc Surg, 9, pp. 437-447Beiguelman, B., Curso prático de bioestatística (1996) Ribeirão Preto: Rev Bras Gen, , 4 a edWilmink, T.B., Quick, C.R., Day, N.E., The association between cigarette smoking and abdominal aortic aneurysm (1999) J Vasc Surg, 30, pp. 1099-1105Heather, B.P., Poskitt, K.R., Earnshaw, J.J., Whyman, M., Shaw, E., Population screening reduces mortality rate aortic aneurysm in men (2000) Br J Surg, 87, pp. 750-753Scott, R.A., Vardulaki, K.A., Walker, N.M., Day, N.E., Duffy, S.W., Ashton, H.A., The long-term benefits of a single scan for abdominal aortic aneurysm (AAA) at age 65 (2001) Eur J Vasc Endovasc Surg, 21, pp. 535-540Johnston, K.W., Rutherford, R.B., Tilson, M.D., Shah, D.M., Hollier, L., Stanley, J.C., Subcommittee on reporting standards for arterial aneurysms. Suggested standards for reporting on arterial aneurysms (1991) J Vasc Surg, 13, pp. 452-458Brown, P.M., Pattenden, R., Gutelius, J.R., The selective management of small abdominal aortic aneurysms: The Kingston study (1992) J Vasc Surg, 15, pp. 21-25. , discussion 25-7Katz, D.J., Stanley, J.C., Zelenock, G.B., Operative mortality rates for intact and ruptured abdominal aortic aneurysms in Michigan: An eleven-year statewide experience (1994) J Vasc Surg, 19, pp. 804-815. , discussion 816-7Cronenwett, J.L., Johnston, K.W., The United Kingdom Small Aneurysm Trial: Implications for surgical treatment of abdominal aortic aneurysms (1999) J Vasc Surg, 29, pp. 191-194Darling, R.C., Messina, C.R., Brewster, D.C., Ottinger, L.W., Autopsy study of unoperated abdominal aortic aneurysm. The case for early resection (1977) Circulation, 56 (3 SUPPL.), pp. II161-II164Sterpetti, A.V., Cavallaro, A., Cavallari, N., Factors influencing the rupture of abdominal aortic aneurysms (1991) Surg Gynecol Obstet, 173, pp. 175-178Bengtsson, H., Bergqvist, D., Ruptured abdominal aortic aneurysm: A population-based study (1993) J Vasc Surg, 18, pp. 74-80Silva, E.S., Dói, A., Hanaoka, B.Y., Takeda, F.R., Ikeda, M.H., Prevalência de aneurismas e outras anormalidades do diâmetro da aorta infra-renal detectadas em necropsia (2002) J Vasc Bras, 1, pp. 89-96Cronenwett, J.L., Sargent, S.K., Wall, M.H., Variables that affect the expansion rate and outcome of small abdominal aortic aneurysms (1990) J Vasc Surg, 11, pp. 260-268. , discussion 268-9Brown, P.M., Pattenden, R., Vernooy, C., Zelt, D.T., Gutelius, J.R., Selective management of abdominal aortic aneurysms in a prospective measurement program (1996) J Vasc Surg, 23, pp. 213-220. , discussion 221-2Irvine, C.D., Shaw, E., Poskitt, K.R., Whyman, M.R., Earnshaw, J.J., Heather, B.P., A comparison of the mortality rate after elective repair of aortic aneurysms detected either by screening or incidentally (2000) Eur J Vasc Endovasc Surg, 20, pp. 374-378Hallin, A., Bergqvist, D., Holmberg, L., Literature review of surgical management of abdominal aortic aneurysm (2001) Eur J Vasc Endovasc Surg, 22, pp. 197-204Scott, R.A.P., Ashton, H.A., Sutton, G.L.J., Ultrasound screening of a general practice for abdominal aortic aneurysm (1986) Br J Surg, 73, p. 318O'Kelly, T.J., Heather, B.P., General practice-based population screening for abdominal aortic aneurysms: A pilot study (1989) Br J Surg, 76, pp. 479-480Scott, R.A., Ashton, H.A., Kay, D.N., Abdominal aortic aneurysm 4237 screened patients: Prevalence, development and management over 6 years (1991) Br J Surg, 78, pp. 1122-1125Lucarotti, M.E., Shaw, E., Heather, B.P., Distribution of aortic diameter in a screened male population (1992) Br J Surg, 79, pp. 641-642Bonamigo TP, Araújo FL, Siqueira I, Becker M. 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Ki-1/57 And Cgi-55 Ectopic Expression Impact Cellular Pathways Involved In Proliferation And Stress Response Regulation

Ki-1/57 (HABP4) and CGI-55 (SERBP1) are regulatory proteins and paralogs with 40.7% amino acid sequence identity and 67.4% similarity. Functionally, they have been implicated in the regulation of gene expression on both the transcriptional and mRNA metabolism levels. A link with tumorigenesis is suggested, since both paralogs show altered expression levels in tumor cells and the Ki-1/57 gene is found in a region of chromosome 9q that represents a haplotype for familiar colon cancer. However, the target genes regulated by Ki-1/57 and CGI-55 are unknown. Here, we analyzed the alterations of the global transcriptome profile after Ki-1/57 or CGI-55 overexpression in HEK293T cells by DNA microchip technology. We were able to identify 363 or 190 down-regulated and 50 or 27 up-regulated genes for Ki-1/57 and CGI-55, respectively, of which 20 were shared between both proteins. Expression levels of selected genes were confirmed by qRT-PCR both after protein overexpression and siRNA knockdown. The majority of the genes with altered expression were associated to proliferation, apoptosis and cell cycle control processes, prompting us to further explore these contexts experimentally. We observed that overexpression of Ki-1/57 or CGI-55 results in reduced cell proliferation, mainly due to a G1 phase arrest, whereas siRNA knockdown of CGI-55 caused an increase in proliferation. In the case of Ki-1/57 overexpression, we found protection from apoptosis after treatment with the ER-stress inducer thapsigargin. Together, our data give important new insights that may help to explain these proteins putative involvement in tumorigenic events.18431229442956Kobarg, J., Schnittger, S., Fonatsch, C., Lemke, H., Bowen, M.A., Buck, F., Hansen, H.P., Characterization, mapping and partial cDNA sequence of the 57-kD intracellular Ki-1 antigen (1997) Exp. Clin. Immunogenet., 14, pp. 273-280Serce, N.B., Boesl, A., Klaman, I., von Serényi, S., Noetzel, E., Press, M.F., Dimmler, A., Dahl, E., Overexpression of SERBP1 (plasminogen activator inhibitor 1 RNA binding protein) in human breast cancer is correlated with favourable prognosis (2012) BMC Cancer, 12, p. 597Heaton, J.H., Dlakic, W.M., Dlakic, M., Gelehrter, T.D., Identification and cDNA cloning of a novel RNA-binding protein that interacts with the cyclic nucleotide-responsive sequence in the type-1 plasminogen activator inhibitor mRNA (2001) J. Biol. Chem., 276, pp. 3341-3347Lemos, T.A., Passos, D.O., Nery, F.C., Kobarg, J., Characterization of a new family of proteins that interact with the C-terminal region of the chromatin-remodeling factor CHD-3 (2003) FEBS Lett., 533, pp. 14-20Kobarg, C.B., Kobarg, J., Crosara-Alberto, D.P., Theizen, T.H., Franchini, K.G., MEF2C DNA-binding activity is inhibited through its interaction with the regulatory protein Ki-1/57 (2005) FEBS Lett., 579, pp. 2615-2622Nery, F.C., Rui, E., Kuniyoshi, T.M., Kobarg, J., Evidence for the interaction of the regulatory protein Ki-1/57 with p53 and its interacting proteins (2006) Biochem. Biophys. Res. Commun., 341, pp. 847-855Bressan, G.C., Quaresma, A.J.C., Moraes, E.C., Manfiolli, A.O., Passos, D.O., Gomes, M.D., Kobarg, J., Functional association of human Ki-1/57 with pre-mRNA splicing events (2009) FEBS J., 276, pp. 3770-3783Gonçalves, K.D.A., Bressan, G.C., Saito, A., Morello, L.G., Zanchin, N.I.T., Kobarg, J., Evidence for the association of the human regulatory protein Ki-1/57 with the translational machinery (2011) FEBS Lett., 585, pp. 2556-2560Nery, F.C., Passos, D.O., Garcia, V.S., Kobarg, J., Ki-1/57 interacts with RACK1 and is a substrate for the phosphorylation by phorbol 12-myristate 13-acetate-activated protein kinase C (2004) J. Biol. Chem., 279, pp. 11444-11455Passos, D.O., Bressan, G.C., Nery, F.C., Kobarg, J., Ki-1/57 interacts with PRMT1 and is a substrate for arginine methylation (2006) FEBS J., 273, pp. 3946-3961Lemos, T.A., Kobarg, J., CGI-55 interacts with nuclear proteins and co-localizes to p80-coilin positive-coiled bodies in the nucleus (2006) Cell Biochem. 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Human Nek7-interactor Rgs2 Is Required For Mitotic Spindle Organization

The mitotic spindle apparatus is composed of microtubule (MT) networks attached to kinetochores organized from 2 centrosomes (a.k.a. spindle poles). In addition to this central spindle apparatus, astral MTs assemble at the mitotic spindle pole and attach to the cell cortex to ensure appropriate spindle orientation. We propose that cell cycle-related kinase, Nek7, and its novel interacting protein RGS2, are involved in mitosis regulation and spindle formation. We found that RGS2 localizes to the mitotic spindle in a Nek7-dependent manner, and along with Nek7 contributes to spindle morphology and mitotic spindle pole integrity. RGS2-depletion leads to a mitotic-delay and severe defects in the chromosomes alignment and congression. Importantly, RGS2 or Nek7 depletion or even overexpression of wild-type or kinase-dead Nek7, reduced γ-tubulin from the mitotic spindle poles. In addition to causing a mitotic delay, RGS2 depletion induced mitotic spindle misorientation coinciding with astral MT-reduction. We propose that these phenotypes directly contribute to a failure in mitotic spindle alignment to the substratum. In conclusion, we suggest a molecular mechanism whereupon Nek7 and RGS2 may act cooperatively to ensure proper mitotic spindle organization.144656667Goshima, G., Wollman, R., Goodwin, S.S., Zhang, N., Scholey, J.M., Vale, R.D., Stuurman, N., Genes required for mitotic spindle assembly in Drosophila S2 cells (2007) Science, 5823, pp. 417-421. , http://dx.doi.org/10.1126/science.1141314Uehara, R., Nozawa, R.S., Tomioka, A., Petry, S., Vale, R.D., Obuse, C., Goshima, G., The augmin complex plays a critical role in spindle microtubule generation for mitotic progression and cytokinesis in human cells (2009) Proc Natl Acad Sci USA, 106, pp. 6998-7003. , http://dx.doi.org/10.1073/pnas.0901587106, PMID:19369198Dumont, S., Mitchison, T.J., Force and length in the mitotic spindle (2009) Curr Biol, 17, pp. R749-R761. , http://dx.doi.org/10.1016/j.cub.2009.07.028Hayward, D., Metz, J., Pellacani, C., Wakefield, J.G., Synergy between multiple microtubule-generating pathways confers robustness to centrosome-driven mitotic spindle formation (2014) Dev Cell, 1, pp. 81-93. , http://dx.doi.org/10.1016/j.devcel.2013.12.001Luders, J., Stearns, T., Microtubule-organizing centres: A re-evaluation (2007) Nat Rev Mol Cell Biol, 8, pp. 161-167. , http://dx.doi.org/10.1038/nrm2100, PMID:17245416Willard, F.S., Kimple, R.J., Siderovski, D.P., Return of the GDI: The GoLoco motif in cell division (2004) Annu Rev Biochem, 73, pp. 925-951. , http://dx.doi.org/10.1146/annurev.biochem.73.011303.073756, PMID:15189163Kotak, S., Gönczy, P., Mechanisms of spindle positioning: Cortical force generators in the limelight (2013) Curr Opin Cell Biol, 6, pp. 741-748. , http://dx.doi.org/10.1016/j.ceb.2013.07.008Zheng, Z., Wan, Q., Liu, J., Zhu, H., Chu, X., Du, Q., Evidence for dynein and astral microtubule-mediated cortical release and transport of Gai/LGN/NuMA complex in mitotic cells (2013) Mol Biol Cell, 7, pp. 901-913. , http://dx.doi.org/10.1091/mbc.E12-06-0458Musacchio, A., Salmon, E.D., The spindle-assembly checkpoint in space and time (2007) Nat Rev Mol Cell Biol, 8, pp. 379-393. , http://dx.doi.org/10.1038/nrm2163, PMID:17426725Noatynska, A., Gotta, M., Meraldi, P., Mitotic spindle (DIS) orientation and DISease: Cause or consequence? 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Characterization Of The Human Nek7 Interactome Suggests Catalytic And Regulatory Properties Distinct From Those Of Nek6

Human NEK7 is a regulator of cell division and plays an important role in growth and survival of mammalian cells. Human NEK6 and NEK7 are closely related, consisting of a conserved C-terminal catalytic domain and a nonconserved and disordered N-terminal regulatory domain, crucial to mediate the interactions with their respective proteins. Here, in order to better understand NEK7 cellular functions, we characterize the NEK7 interactome by two screening approaches: one using a yeast two-hybrid system and the other based on immunoprecipitation followed by mass spectrometry analysis. These approaches led to the identification of 61 NEK7 interactors that contribute to a variety of biological processes, including cell division. Combining additional interaction and phosphorylation assays from yeast two-hybrid screens, we validated CC2D1A, TUBB2B, MNAT1, and NEK9 proteins as potential NEK7 interactors and substrates. Notably, endogenous RGS2, TUBB, MNAT1, NEK9, and PLEKHA8 localized with NEK7 at key sites throughout the cell cycle, especially during mitosis and cytokinesis. Furthermore, we obtained evidence that the closely related kinases NEK6 and NEK7 do not share common interactors, with the exception of NEK9, and display different modes of protein interaction, depending on their N- and C-terminal regions, in distinct fashions. In summary, our work shows for the first time a comprehensive NEK7 interactome that, combined with functional in vitro and in vivo assays, suggests that NEK7 is a multifunctional kinase acting in different cellular processes in concert with cell division signaling and independently of NEK6.13940744090Ma, H.T., Poon, R.Y., How protein kinases co-ordinate mitosis in animal cells (2011) Biochem. 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