William Brown

Contact

• workRoom D08 Medical School
  Queen's Medical Centre
  Nottingham
  NG7 2UH
  UK
• work0115 823 0386
• fax0115 823 0338
• william.brown@nottingham.ac.uk

Biography

BA Nat.Sci. Class1: (Biochemistry), Cambridge; 1976, D.Phil. Oxford: 1981, Reader in Genetics: Nottingham University: 2000- present

Research Summary

The genetics and evolution of centromere assembly in vertebrates and fission yeast: We are using a set of engineered mini-chromosomes to investigate the genetics and evolution of centromere assembly… read more

Selected Publications

• RHIND, N., CHEN, Z., YASSOUR, M., THOMPSON, D.A., HAAS, B.J., HABIB, N., WAPINSKI, I., ROY, S., LIN, M.F., HEIMAN, D.I., YOUNG, S.K., FURUYA, K., GUO, Y., PIDOUX, A. and CHEN, H.M., 2011. Comparative functional genomics of the fission yeasts Science (New York, N.Y.). 332(6032), 930-936
• BROWN, W.R.A., LITI, G., ROSA, C., JAMES, S., ROBERTS, I., ROBERT, V., JOLLY, N., TANG, W., BAUMANN, P., GREEN, C., SCHLEGEL, K., YOUNG, J., HIRCHAUD, F., LEEK, S., THOMAS, G., BLOMBERG, A. and WARRINGER, J., 2011. A geographically diverse collection of Schizosaccharomyces pombe isolates shows limited phenotypic variation but extensive karyotypic diversity G3: Genes, Genomes, Genetics. 1(7), 615-626
• BROOKFIELD, R., DAFHNIS-CALAS, F., XU, Z. and BROWN, W., 2012. Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of "Rong And Golic" Transgenic Research: Springer. 21(5), 1125-1136
• DODGSON, J., BROWN, W., ROSA, C.A. and ARMSTRONG, J., 2010. Reorganization of the growth pattern of Schizosaccharomyces pombe in invasive filament formation Eukaryotic Cell. 9(11), 1788-1797

• View all publications

.txt Files

• ApraPhiC31attPneopBAC3e.txt
• BloxPBsrBmod.txt
• CCAG(5'NLS)phiC31integraseIRESZeo.txt
• CCAGCREIRESGPT.txt
• CCAGCREIRESNEO.txt
• CCAGloxPHytkP.txt
• phiC31attP Neo.txt
• phiC31BACBCCAGloxPBsrBmod.txt
• PhiC31BACBloxPBsrBmod.txt
• phiC31BACPCCAGloxPHytkP.txt
• phiC31PACBloxPBsrBmod.txt
• phiC31PACPloxPHytkP.txt
• PloxPHytkP.txt

.gb Files

• ApraPhiC31attPneopBAC3e.gb
• BloxPBsrBmod.gb
• CCAG(5'NLS)phiC31integraseIRESZeo.gb
• CCAGCREIRESNEO.gb
• CCAGCREIRESNEO.gb
• CCAGloxPHytkP.gb
• phiC31attP Neo.gb
• phiC31BACBCCAGloxPBsrBmod.gb
• PhiC31BACBloxPBsrBmod.gb
• phiC31BACPCCAGloxPHytkP.gb
• phiC31PACBloxPBsrBmod.gb
• phiC31PACPloxPhytkP.gb
• PloxPHytkP.gb

Current Research

The genetics and evolution of centromere assembly in vertebrates and fission yeast: We are using a set of engineered mini-chromosomes to investigate the genetics and evolution of centromere assembly in vertebrates and fission yeast. Our work has led us to develop several new techniques for chromosome engineering and we are also applying these to the construction of a plant artificial chromosome.

Human mini-chromosome engineering

Centromeres mediate the segregation of chromosomes at cell division; a function that is conserved throughout eucaryotic evolution. However centromeric DNA is amongst the most rapidly evolving in the genome and thus poses a paradox which we should like to resolve. In order to do this we need to understand how centromeric DNA is recognized. We are using vertebrate cells and fission yeast to address this question. In both organisms centromeres are comprised of long stretches of DNA. These sequences are hard to manipulate experimentally and so much of our effort has been devoted to developing methods for manipulating such long tracts of DNA. In our early work we used telomeres as in vivo reagents to fragment human chromosomes. This worked well and generated a set of mini-chromosomes that have been useful in subsequent experiments and told us that the minimum size for a centromere in human cells was about 100kb. More recently we have developed a new type of site specific recombinase for use in both vertebrate cells. At the moment we are using this type of enzyme to swap the existing centromere sequence on a yeast or human mini-chromosome with a defined tract of sequence whose function we wish to test.

Our technology for manipulating chromosome structure is of general applicability and we are also using it to build artificial plant chromosomes. Our approach is described below.

Engineering the Human Y chromosome by telomere directed chromosome breakage

Chromosome engineering and centromere function

Sunir Malla PhD student, Felix Dafnis postdoc and William Brown

Outline: We are developing new methods for engineering chromosome structure and using them to study the mechanism and sequence specificity of centromere assembly in chicken and human cells.

Details:

1. We are establishing a method for assembling very large (100kb -1Mb) tracts of transgenic DNA on vertebrate chromosomes

2. We are establishing a method for irreversibly deleting DNA sequences of arbitrary length from human chromosomes.

3. We are using these methods to engineer human and other mini-chromosomes with centromeric DNA of defined length and sequence and then studying how these different centromeres work in chicken, mouse and human cells.

Support: EU

Chick

Human

Mouse

Chromosome engineering and mouse centromere function

Felix Dafnis

Outline: I am studying the mechanism and sequence specificity of centromere assembly in mouse cells.

Details:

1. In collaboration with William and Sunir I am establishing a method for assembling very large (100kb -1Mb) tracts of transgenic DNA on vertebrate chromosomes

2. I am using this method to build centromeres on mini-chromosomes that are unstable in mouse cells with the object of identifying the features of the sequence necessary for centromere function in mouse cells.

Support: EU

Chromosome engineering and yeast centromere function

Engineering a rice artificial chromosome

Zhang Yao Xu, Leverhulme Trust funded post-doc

Outline: I am using the methods of chromosome engineering established in our group to convert a human mini-chromosome into a plant artificial chromosome. The long term goal is to develop an artificial chromosome vector for this important crop plant.

Details:

I am taking one of William's mini-chromosomes and adding to it a variety of cis-acting sequences that will enable it to function as a chromosome in plant cells. These sequences include, of course, centromeric DNA, a selectable marker gene and telomeres. I will then introduce the modified mini-chromosome into plant protoplasts by somatic cell fusion techniques. This work is carried out in collaboration with Plant Sciences at the Sutton Bonnington Campus.

©Robert Soreng. Courtesy of Smithsonian Institution, Department of Systematic Biology-Botany. Usage Guidelines.

• XU, Z and BROWN, W.R.A, 2016. Comparison and optimization of ten phage encoded serine integrases for genome engineering in Saccharomyces cerevisiae. BMC Biotechnol.. 16(1), 13
• JEFFARES DC, RALLIS C, RIEUX A, SPEED D, PŘEVOROVSKÝ M, MOURIER T, MARSELLACH FX, IQBAL Z, LAU W, CHENG TM, PRACANA R, MÜLLEDER M, LAWSON JL, CHESSEL A, BALA S, HELLENTHAL G, O'FALLON B, KEANE T, SIMPSON JT, BISCHOF L, TOMICZEK B, BITTON DA, SIDERI T, CODLIN S, HELLBERG JE, VAN TRIGT L, JEFFERY L, LI JJ, ATKINSON S, THODBERG M, FEBRER M, MCLAY K, DROU N, BROWN W, HAYLES J, CARAZO SALAS RE, RALSER M, MANIATIS N, BALDING DJ, BALLOUX F, DURBIN R and BÄHLER J, 2015. The genomic and phenotypic diversity of Schizosaccharomyces pombe. Nature genetics. 47(3), 235-41
• BROWN WRA, THOMAS G, LEE NCO, BLYTHE M, LITI G, WARRINGER J and LOOSE MW, 2014. Kinetochore assembly and heterochromatin formation occur autonomously in Schizosaccharomyces pombe. Proceedings of the National Academy of Sciences of the United States of America. 111(5), 1903-8
• XU, Z., THOMAS, L., DAVIES, B., CHALMERS, R., SMITH, M. and BROWN, W., 2013. Accuracy and efficiency define Bxb1 integrase as the best of fifteen candidate serine recombinases for the integration of DNA into the human genome BMC Biotechnology. 13, 87
• EARNSHAW, W.C, ALLSHIRE, R.C, BLACK, B.E, BLOOM, K, BRINKLEY, B.R, BROWN, W, CHEESMAN, I.M, CHOO, K.H.A, COPENHAVER, G.P, DELUCA, J.G, DESAI, A, DIEKMANN, S, ERHARDT, S, FITZGERALD-HAYES, M, FOLTZ, D, FUKAGAWA, T, GASSMANN, R, GERLICH, D.W, GLOVER, D.M, GORBSKY, G.J, HARRISON, S.C, HEUN, P, HIROTA, T, JANSEN, L.E.T, KARPEN, G, KOPS, G.J.P.L, LAMPSON, M.A., LENS, S.M, LOSADA, A, LUGER, K, MAIATO, H, MADDOX, P.S, MARGOLIS, R.L, MASUMOTO H, MCAINSH, A.D, MELLONE, B.G, MERALDI, P, MUSACCHIO, A, OEGEMA, K, O'NEILL, R.J, SALMON, E.D, SCOTT, K.C, STRAIGHT, A.F, STUKENBERG, P.T, SULLIVAN, B.A, SULLIVAN, K.F, SUNKEL, C.E, SWEDLOW, J.R, WALCZAK, C.E, WARBURTON, P.E, WESTERMANN, S, WILLARD, H.F, WORDEMAN, L, YANAGIDA, M, YEN, T.J, YODA, K and CLEVELAND, D.W, 2013. Esperanto for histones: CENP-A, not CenH3, is the centromeric histone H3 variant Chromosome Research. 21(2), 101-106
• BROOKFIELD, R., DAFHNIS-CALAS, F., XU, Z. and BROWN, W., 2012. Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of "Rong And Golic" Transgenic Research: Springer. 21(5), 1125-1136
• RHIND, N., CHEN, Z., YASSOUR, M., THOMPSON, D.A., HAAS, B.J., HABIB, N., WAPINSKI, I., ROY, S., LIN, M.F., HEIMAN, D.I., YOUNG, S.K., FURUYA, K., GUO, Y., PIDOUX, A. and CHEN, H.M., 2011. Comparative functional genomics of the fission yeasts Science (New York, N.Y.). 332(6032), 930-936
• BROWN, W.R.A., LEE, N.C.O., XU, Z. and SMITH, M.C.M., 2011. Serine recombinases as tools for genome engineering Methods. 53(4), 372-379
• BROWN, W.R.A., LITI, G., ROSA, C., JAMES, S., ROBERTS, I., ROBERT, V., JOLLY, N., TANG, W., BAUMANN, P., GREEN, C., SCHLEGEL, K., YOUNG, J., HIRCHAUD, F., LEEK, S., THOMAS, G., BLOMBERG, A. and WARRINGER, J., 2011. A geographically diverse collection of Schizosaccharomyces pombe isolates shows limited phenotypic variation but extensive karyotypic diversity G3: Genes, Genomes, Genetics. 1(7), 615-626
• DODGSON, J., BROWN, W., ROSA, C.A. and ARMSTRONG, J., 2010. Reorganization of the growth pattern of Schizosaccharomyces pombe in invasive filament formation Eukaryotic Cell. 9(11), 1788-1797
• SMITH, M.C.M, BROWN, W.R.A, MCEWAN, A.R and ROWLEY, P.A, 2010. Site-specific recombination by φC31 integrase and other large serine recombinases Biochemical Society Transaction. 38(2), 388-394
• BROWN, WILLIAM R. A. and XU, ZHENG-YAO, 2009. The 'kinetochore maintenance loop' - The mark of regulation? BioEssays. 31(2), 228-236
• XU, Z, LEE, N. C and DAFHNIS-CALAS, F. MALLA, S. SMITH, M. C.M AND BROWN, W. R.A, 2007. Site-specific recombination in Schizosaccharomyces pombe and systematic assembly of a 400kb transgene array in mammalian cells using the integrase of Streptomyces 'phage phiBT1 Nucleic Acids Research. 36(1), Epub 2007 Dec 20
• BROWN, WILLIAM R A, SMITH, MARGARET C M, DAFHNIS-CALAS, FELIX, MALLA, SUNIR and XU, ZHENGYAO, 2006. Chromosome engineering in DT40 cells and mammalian centromere function. Sub-cellular biochemistry. 40, 39-48
• MALLA, S., DAFHNIS-CALAS, F., BROOKFIELD, J. F., SMITH, M. C. and BROWN, W. R., 2005. Rearranging the centromere of the human Y chromosome with φC31 integrase Nucleic Acids Research. 33(19), 6101-6113
• DAFHNIS-CALAS, F., XU, Z., HAINES, S., MALLA, S.K., SMITH, M.C.M. and BROWN, W.R.A., 2005. Iterative in vivo assembly of large and complex transgenes by combining the activities of φC31 integrase and Cre recombinase. Nucleic Acids Research. 33(22), e189
• RÉGNIER, V., VAGNARELLI, P., FUKAGAWA, T., ZERJAL, T., BURNS, E., TROUCHE, D., EARNSHAW, W. and BROWN, W., 2005. CENP-A is required for accurate chromosome segregation and sustained kinetochore association of BubR1 Molecular and Cellular Biology. 25(10), 3967-3981
• HOLLAND, S., IOANNOU, D., HAINES, S. and BROWN, W.R.A., 2005. Comparison of Dam tagging and chromatin immunoprecipitation as tools for the identification of the binding sites for S. pombe CENP-C. Chromosome Research. 13(1), 73-83
• HUBBARD, S. J., GRAFHAM, D. V., BEATTIE, K. J., OVERTON, I. M., MCLAREN, S. R., CRONING, M. D. R., BOARDMAN, P. E., BONFIELD, J. K., BURNSIDE, J., DAVIES, R. M, FARRELL, ER, FRANCIS, MD, GRIFFITH-JONES, S, HUMPHRAY, SJ, SCOTT, CE, TANG, H, TAYLOR, RG, TICKLE, C, BROWN, WR, BIRNEY, E and ROGERS,J>, WILSON, SA, 2005. Transcriptome analysis for the chicken based on 19,626 finished cDNA sequences and 485,337 expressed sequence tags Genome Research. VOL 15(NUMB 1), 174-183
• BROWN, W. R. A., HUBBARD, S. J., TICKLE, C. and WILSON, S. A., 2003. The chicken as a model for large-scale analysis of vertebrate gene function Nature Reviews: Genetics. VOL 4(PART 2), 87-98
• MEE, P. J., SHEN, M. H., SMITH, A. G. and BROWN, W. R. A., 2003. An unpaired mouse centromere passes consistently through male meiosis and does not significantly compromise spermatogenesis Chromosoma. VOL 112(PART 4), 183-189
• KOUPRINA, N., EBERSOLE, T., KORIABINE, M., PAK, E., ROGOZIN, I. B., KATOH, M., OSHIMURA, M., OGI, K., PEREDELCHUK, M. and SOLOMON, G., 2003. Cloning of human centromeres by transformation-associated recombination in yeast and generation of functional human artificial chromosomes Nucleic Acids Research. VOL 31(PART 3), 922-934
• RÉGNIER, V., NOVELLI, J., FUKAGAWA, T., VAGNARELLI, P. and BROWN, W., 2003. Characterization of chicken CENP-A and comparative sequence analysis of vertebrate centromere-specific histone H3-like proteins Gene. 316, 39-46
• BOARDMAN, P.E., SANZ-EZQUERRO, J., OVERTON, I.M., BURT, D.W., BOSCH, E., FONG, W.T., TICKLE, C., BROWN, W.R.A, WILSON, S.A. and HUBBARD, S.J., 2002. A comprehensive collection of chicken cDNAs Current Biology. 12(22), 1965-1969
• FUKAGAWA, T., MIKAMI, Y., NISHIHASHI, A., REGNIER, V., HARAGUCHI, T., HIRAOKA, Y., SUGATA, N., TODOKORO, K., BROWN, W. and IKEMURA, T., 2001. CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells EMBO Journal. 20(16), 4603-4617
• SHEN, M. H., YANG, J. W., YANG, J., PENDON, C. and BROWN, W. R., 2000. The accuracy of segregation of human mini-chromosomes varies in different vertebrate cell lines, correlates with the extent of centromere formation and provides evidence for a trans-acting centromere maintenance activity Chromosoma. VOL 109(PART 8), 524-535
• YANG, J. W., PENDON, C., YANG, J., HAYWOOD, N., CHAND, A. and BROWN, W. R. A., 2000. Human mini-chromosomes with minimal centromeres Human Molecular Genetics. VOL 9(PART 12), 1891-1902
• BROWN, W.R.A, MEE, P.J and HONG SHEN, M, 2000. Artificial chromosomes: Ideal vectors? Trends in Biotechnology. 18(5), 218-223
• SHEN, M.H, MEE, P.J, NICHOLAS, J, YANG, J, BROOK, F, GARDNER, R.L, SMITH, A.G and BROWN, W.R.A, 2000. A structurally defined mini-chromosome vector for the mouse germ line Current Biology. 10(1), 31-34
• FUKAGAWA, T, HAYWARD, N, YANG, JA, AZZALIN, C, GRIFFIN, D, STEWART, AF and BROWN, W, 1999. The chicken HPRT gene: a counter selectable marker for the DT40 cell line NUCLEIC ACIDS RESEARCH. 27(9), 1966-1969
• FUKAGAWA, T. and BROWN, W. R. A., 1997. Efficient conditional mutation of the vertebrate CENP-C gene Human Molecular Genetics. VOL 6(NUMBER 13), 230
• SHEN, MH, YANG, J, LOUPART, ML, SMITH, A and BROWN, W, 1997. Human mini-chromosomes in mouse embryonal stem cells HUMAN MOLECULAR GENETICS. 6(8), 1375-1382
• HELLER, R., BROWN, K. E., BURGTORF, C. and BROWN, W. R. A., 1996. Mini-chromosomes derived from the human Y chromosome by telomere directed chromosome breakage Proceedings of the National Academy of Sciences of the United States of America. VOL 93(NUMBER 14), 7125-7130
• NING, Y, ROSCHKE, A, SMITH, ACM, MACHA, M, PRECHT, K, RIETHMAN, H, LEDBETTER, DH, FLINT, J, HORSLEY, S, REGAN, R, KEARNEY, L, KNIGHT, S, KVALOY, K and BROWN, WRA, 1996. A complete set of human telomeric probes and their clinical application NATURE GENETICS. 14(1), 86-89
• BROWN W, HELLER R, LOUPART ML, SHEN MH and CHAND A, 1996. Mammalian artificial chromosomes. Current opinion in genetics & development. 6(3), 281-8
• BROWN W and TYLER-SMITH C, 1995. Centromere activation. Trends in genetics : TIG. 11(9), 337-9
• BROWN, KE, BARNETT, MA, BURGTORF, C, SHAW, P, BUCKLE, VJ and BROWN, WRA, 1994. DISSECTING THE CENTROMERE OF THE HUMAN Y-CHROMOSOME WITH CLONED TELOMERIC DNA HUMAN MOLECULAR GENETICS. 3(8), 1227-1237
• KVALOY, K, GALVAGNI, F and BROWN, WRA, 1994. THE SEQUENCE ORGANIZATION OF THE LONG ARM PSEUDOAUTOSOMAL REGION OF THE HUMAN SEX-CHROMOSOMES HUMAN MOLECULAR GENETICS. 3(5), 771-778
• BARNETT, MA, BUCKLE, VJ, EVANS, EP, PORTER, ACG, ROUT, D, SMITH, AG and BROWN, WRA, 1993. TELOMERE DIRECTED FRAGMENTATION OF MAMMALIAN CHROMOSOMES NUCLEIC ACIDS RESEARCH. 21(1), 27-36
• BROWN W R A, 1992. TELOMERASE AND CHROMOSOME HEALING Current Biology. 2(3), 127-129
• ITZHAKI, JE, BARNETT, MA, MACCARTHY, AB, BUCKLE, VJ, BROWN, WRA and PORTER, ACG, 1992. TARGETED BREAKAGE OF A HUMAN-CHROMOSOME MEDIATED BY CLONED HUMAN TELOMERIC DNA NATURE GENETICS. 2(4), 283-287
• WILKIE, AOM, HIGGS, DR, RACK, KA, BUCKLE, VJ, SPURR, NK, FISCHELGHODSIAN, N, CECCHERINI, I, BROWN, WRA and HARRIS, PC, 1991. STABLE LENGTH POLYMORPHISM OF UP TO 260 KB AT THE TIP OF THE SHORT ARM OF HUMAN CHROMOSOME-16 CELL. 64(3), 595-606
• BROWN, WRA, MACKINNON, PJ, VILLASANTE, A, SPURR, N, BUCKLE, VJ and DOBSON, MJ, 1990. STRUCTURE AND POLYMORPHISM OF HUMAN TELOMERE-ASSOCIATED DNA CELL. 63(1), 119-132
• BROWN, WRA, DOBSON, MJ and MACKINNON, P, 1990. TELOMERE CLONING AND MAMMALIAN CHROMOSOME ANALYSIS JOURNAL OF CELL SCIENCE. 95, 521-526
• BROWN, WRA, 1989. MOLECULAR-CLONING OF HUMAN TELOMERES IN YEAST NATURE. 338(6218), 774-776
• BROWN, WRA, 1988. A PHYSICAL MAP OF THE HUMAN PSEUDOAUTOSOMAL REGION EMBO JOURNAL. 7(8), 2377-2385
• SOUTHERN, EM, ANAND, R, BROWN, WRA and FLETCHER, DS, 1987. A MODEL FOR THE SEPARATION OF LARGE DNA-MOLECULES BY CROSSED-FIELD GEL-ELECTROPHORESIS NUCLEIC ACIDS RESEARCH. 15(15), 5925-5943
• TYLERSMITH, C and BROWN, WRA, 1987. STRUCTURE OF THE MAJOR BLOCK OF ALPHOID SATELLITE DNA ON THE HUMAN Y-CHROMOSOME JOURNAL OF MOLECULAR BIOLOGY. 195(3), 457-470
• BROWN, WRA and BIRD, AP, 1986. LONG-RANGE RESTRICTION SITE MAPPING OF MAMMALIAN GENOMIC DNA NATURE. 322(6078), 477-481
• COOKE, HJ, BROWN, WRA and RAPPOLD, GA, 1985. HYPERVARIABLE TELOMERIC SEQUENCES FROM THE HUMAN SEX-CHROMOSOMES ARE PSEUDOAUTOSOMAL NATURE. 317(6039), 687-692