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9. Double diffusion as a driver of turbulence in the stratified boundary layer beneath George VI Ice Shelf

Author

Middleton, L., Davis, P.E.D., Taylor, J.R., Nicholls, K.W., Middleton, L., Davis, P.E.D., Taylor, J.R., and Nicholls, K.W.

Abstract

Warmer and more persistent intrusions of Circumpolar Deep Water (CDW) onto the West Antarctic Peninsula are a key driver of the recent increase in ice shelf mass loss. The relatively warm and salty CDW is thought to be mixed up to the base of the ice shelves via shear-driven turbulence where it has a high capacity to melt the ice. We analyze data from a year-long mooring beneath George VI Ice Shelf at a location where double-diffusive layering was observed. The turbulent dissipation rates do not vary with mean flow speed, suggesting shear-driven mixing is not the driver of basal melt at this site. Instead, we predict the observed dissipation using a new method that links along-isopycnal stirring of temperature anomalies with double-diffusive convection. Our work suggests that along-isopycnal temperature variance may be a stronger indicator of melt than flow speed within strongly stratified ice shelf-ocean boundary layers.

Published
2022

11. Calibration of the LIGO gravitational wave detectors in the fifth science run

Author

Abadie, J., Abbott, B.P., Abbott, R., Abernathy, M., Adams, C., Adhikari, R., Ajith, P., Allen, B., Allen, G., Amador Ceron, E., Amin, R.S., Anderson, S.B., Anderson, W.G., Arain, M.A., Araya, M., Aronsson, M., Aso, Y., Aston, S., Atkinson, D.E., Aufmuth, P., Aulbert, C., Babak, S., Baker, P., Ballmer, S., Barker, D., Barnum, S., Barr, B., Barriga, P., Barsotti, L., Barton, M.A., Bartos, I., Bassiri, R., Bastarrika, M., Bauchrowitz, J., Behnke, B., Benacquista, M., Bertolini, A., Betzwieser, J., Beveridge, N., Beyersdorf, P.T., Bilenko, I.A., Billingsley, G., Birch, J., Biswas, R., Black, E., Blackburn, J.K., Blackburn, L., Blair, D., Bland, B., Bock, O., Bodiya, T.P., Bondarescu, R., Bork, R., Born, M., Bose, S., Boyle, M., Brady, P.R., Braginsky, V.B., Brau, J.E., Breyer, J., Bridges, D.O., Brinkmann, M., Britzger, M., Brooks, A.F., Brown, D.A., Buonanno, A., Burguet–Castell, J., Burmeister, O., Byer, R.L., Cadonati, L., Cain, J., Camp, J.B., Campsie, P., Cannizzo, J., Cannon, K.C., Cao, J., Capano, C., Caride, S., Caudill, S., Cavaglià, M., Cepeda, C., Chalermsongsak, T., Chalkley, E., Charlton, P., Chelkowski, S., Chen, Y., Christensen, N., Chua, S.S.Y., Chung, C.T.Y., Clark, D., Clark, J., Clayton, J.H., Conte, R., Cook, D., Corbitt, T.R., Cornish, N., Costa, C.A., Coward, D.M., Coyne, D.C., Creighton, J.D.E., Creighton, T.D., Cruise, A.M., Culter, R.M., Cumming, A., Cunningham, L., Dahl, K., Danilishin, S.L., Dannenberg, R., Danzmann, K., Das, K., Daudert, B., Davies, G., Davis, A., Daw, E.J., Dayanga, T., DeBra, D., Degallaix, J., Dergachev, V., DeRosa, R., DeSalvo, R., Devanka, P., Dhurandhar, S., Di Palma, I., Díaz, M., Donovan, F., Dooley, K.L., Doomes, E.E., Dorsher, S., Douglas, E.S.D., Drever, R.W.P., Driggers, J.C., Dueck, J., Dumas, J.-C., Eberle, T., Edgar, M., Edwards, M., Effler, A., Ehrens, P., Engel, R., Etzel, T., Evans, M., Evans, T., Fairhurst, S., Fan, Y., Farr, B.F., Fazi, D., Fehrmann, H., Feldbaum, D., Finn, L.S., Flanigan, M., Flasch, K., Foley, S., Forrest, C., Forsi, E., Fotopoulos, N., Frede, M., Frei, M., Frei, Z., Freise, A., Frey, R., Fricke, T.T., Friedrich, D., Fritschel, P., Frolov, V.V., Fulda, P., Fyffe, M., Garofoli, J.A., Gholami, I., Ghosh, S., Giaime, J.A., Giampanis, S., Giardina, K.D., Gill, C., Goetz, E., Goggin, L.M., González, G., Gorodetsky, M.L., Goßler, S., Graef, C., Grant, A., Gras, S., Gray, C., Greenhalgh, R.J.S., Gretarsson, A.M., Grosso, R., Grote, H., Grunewald, S., Gustafson, E.K., Gustafson, R., Hage, B., Hall, P., Hallam, J.M., Hammer, D., Hammond, G., Hanks, J., Hanna, C., Hanson, J., Harms, J., Harry, G.M., Harry, I.W., Harstad, E.D., Haughian, K., Hayama, K., Hayler, T., Heefner, J., Heng, I.S., Heptonstall, A.W., Hewitson, M., Hild, S., Hirose, E., Hoak, D., Hodge, K.A., Holt, K., Hosken, D.J., Hough, J., Howell, E.J., Hoyland, D., Hughey, B., Husa, S., Huttner, S.H., Huynh–Dinh, T., Ingram, D.R., Inta, R., Isogai, T., Ivanov, A., Johnson, W.W., Jones, D.I., Jones, G., Jones, R., Ju, L., Kalmus, P., Kalogera, V., Kandhasamy, S., Kanner, J.B., Katsavounidis, E., Kawabe, K., Kawamura, S., Kawazoe, F., Kells, W., Keppel, D.G., Khalaidovski, A., Khalili, F.Y., Khazanov, E.A., Kim, H., King, P.J., Kinzel, D.L., Kissel, J.S., Klimenko, S., Kondrashov, V., Kopparapu, R., Koranda, S., Kozak, D., Krause, T., Kringel, V., Krishnamurthy, S., Krishnan, B., Kuehn, G., Kullman, J., Kumar, R., Kwee, P., Landry, M., Lang, M., Lantz, B., Lastzka, N., Lazzarini, A., Leaci, P., Leong, J., Leonor, I., Li, J., Lin, H., Lindquist, P.E., Lockerbie, N.A., Lodhia, D., Lormand, M., Lu, P., Luan, J., Lubinski, M., Lucianetti, A., Lück, H., Lundgren, A., Machenschalk, B., MacInnis, M., Mageswaran, M., Mailand, K., Mak, C., Mandel, I., Mandic, V., Márka, S., Márka, Z., Maros, E., Martin, I.W., Martin, R.M., Marx, J.N., Mason, K., Matichard, F., Matone, L., Matzner, R.A., Mavalvala, N., McCarthy, R., McClelland, D.E., McGuire, S.C., McIntyre, G., McIvor, G., McKechan, D.J.A., Meadors, G., Mehmet, M., Meier, T., Melatos, A., Melissinos, A.C., Mendell, G., Menéndez, D.F., Mercer, R.A., Merill, L., Meshkov, S., Messenger, C., Meyer, M.S., Miao, H., Miller, J., Mino, Y., Mitra, S., Mitrofanov, V.P., Mitselmakher, G., Mittleman, R., Moe, B., Mohanty, S.D., Mohapatra, S.R.P., Moraru, D., Moreno, G., Morioka, T., Mors, K., Mossavi, K., MowLowry, C.M., Mueller, G., Mukherjee, S., Mullavey, A., Müller-Ebhardt, H., Munch, J., Murray, P.G., Nash, T., Nawrodt, R., Nelson, J., Newton, G., Nishizawa, A., Nolting, D., Ochsner, E., O’Dell, J., Ogin, G.H., Oldenburg, R.G., O’Reilly, B., O’Shaughnessy, R., Osthelder, C., Ottaway, D.J., Ottens, R.S., Overmier, H., Owen, B.J., Page, A., Pan, Y., Pankow, C., Papa, M.A., Pareja, M., Patel, P., Pathak, D., Pedraza, M., Pekowsky, L., Penn, S., Peralta, C., Perreca, A., Pickenpack, M., Pinto, I.M., Pitkin, M., Pletsch, H.J., Plissi, M.V., Postiglione, F., Predoi, V., Price, L.R., Prijatelj, M., Principe, M., Prix, R., Prokhorov, L., Puncken, O., Quetschke, V., Raab, F.J., Radke, T., Radkins, H., Raffai, P., Rakhmanov, M., Rankins, B., Raymond, V., Reed, C.M., Reed, T., Reid, S., Reitze, D.H., Riesen, R., Riles, K., Roberts, P., Robertson, N.A., Robinson, C., Robinson, E.L., Roddy, S., Röver, C., Rollins, J., Romano, J.D., Romie, J.H., Rowan, S., Rüdiger, A., Ryan, K., Sakata, S., Sakosky, M., Salemi, F., Sammut, L., Sancho de la Jordana, L., Sandberg, V., Sannibale, V., Santamaría, L., Santostasi, G., Saraf, S., Sathyaprakash, B.S., Sato, S., Satterthwaite, M., Saulson, P.R., Savage, R., Schilling, R., Schnabel, R., Schofield, R.M.S., Schulz, B., Schutz, B.F., Schwinberg, P., Scott, J., Scott, S.M., Searle, A.C., Seifert, F., Sellers, D., Sengupta, A.S., Sergeev, A., Shaddock, D.A., Shapiro, B., Shawhan, P., Shoemaker, D.H., Sibley, A., Siemens, X., Sigg, D., Singer, A., Sintes, A.M., Skelton, G., Slagmolen, B.J.J., Slutsky, J., Smith, J.R., Smith, M.R., Smith, N.D., Somiya, K., Sorazu, B., Speirits, F.C., Stein, A.J., Stein, L.C., Steinlechner, S., Steplewski, S., Stochino, A., Stone, R., Strain, K.A., Strigin, S., Stroeer, A., Stuver, A.L., Summerscales, T.Z., Sung, M., Susmithan, S., Sutton, P.J., Szokoly, G.P., Talukder, D., Tanner, D.B., Tarabrin, S.P., Taylor, J.R., Taylor, R., Thomas, P., Thorne, K.A., Thorne, K.S., Thrane, E., Thüring, A., Titsler, C., Tokmakov, K.V., Torres, C., Torrie, C.I., Traylor, G., Trias, M., Tseng, K., Turner, L., Ugolini, D., Urbanek, K., Vahlbruch, H., Vaishnav, B., Vallisneri, M., Van Den Broeck, C., van der Sluys, M.V., van Veggel, A.A., Vass, S., Vaulin, R., Vecchio, A., Veitch, J., Veitch, P.J., Veltkamp, C., Villar, A.E., Vorvick, C., Vyachanin, S.P., Waldman, S.J., Wallace, L., Wanner, A., Ward, R.L., Wei, P., Weinert, M., Weinstein, A.J., Weiss, R., Wen, L., Wen, S., Wessels, P., West, M., Westphal, T., Wette, K., Whelan, J.T., Whitcomb, S.E., White, D.J., Whiting, B.F., Wilkinson, C., Willems, P.A., Williams, L., Willke, B., Winkelmann, L., Winkler, W., Wipf, C.C., Wiseman, A.G., Woan, G., Wooley, R., Worden, J., Yakushin, I., Yamamoto, H., Yamamoto, K., Yeaton-Massey, D., Yoshida, S., Yu, P.P., Zanolin, M., Zhang, L., Zhang, Z., Zhao, C., Zotov, N., Zucker, M.E., and Zweizig, J.

Published
2010
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15. Contribution of fish to the marine inorganic carbon cycle

Author

Wilson, R.W., Millero, F.J., Taylor, J.R., Walsh, P.J., Christensen, V., Jennings, S., and Grosell, M.

Subjects
Carbon cycle (Biogeochemistry) -- Research, Marine fishes -- Environmental aspects, Marine fishes -- Chemical properties, Science and technology
Abstract

Oceanic production of calcium carbonate is conventionally attributed to marine plankton (coccolithophores and foraminifera). Here we report that marine fish produce precipitated carbonates within their intestines and excrete these at high rates. When combined with estimates of global fish biomass, this suggests that marine fish contribute 3 to 15% of total oceanic carbonate production. Fish carbonates have a higher magnesium content and solubility than traditional sources, yielding faster dissolution with depth. This may explain up to a quarter of the increase in titratable alkalinity within 1000 meters of the ocean surface, a controversial phenomenon that has puzzled oceanographers for decades. We also predict that fish carbonate production may rise in response to future environmental changes in carbon dioxide, and thus become an increasingly important component of the inorganic carbon cycle.

Published
2009

16. Multimodality

Author

Forceville, C., Xu, W., Taylor, J.R., and ACLC (FGw)

Abstract

While it is crystal clear that communication can draw on many semiotic resources, research in the humanities has hitherto strongly focused on its verbal manifestations. “Multimodality” labels a variety of approaches and theories trying to remedy this bias by investigating how for instance visuals, music, and sound contribute to meaning-making. The contours of what is developing into a new discipline begin to be discernible. This chapter provides a brief survey of various perspectives on multimodality, addresses the thorny issue of what should count as a mode, and makes suggestions for further development of the fledgling discipline.

Published
2021

19. A meta-analysis of rectal NSAIDs in the prevention of post-ERCP pancreatitis

Author

Elmunzer, B.J., Waljee, A.K., Elta, G.H., Taylor, J.R., Fehmi, S.M.A., and Higgins, P.D.R.

Subjects
Nonsteroidal anti-inflammatory drugs -- Dosage and administration, Nonsteroidal anti-inflammatory drugs -- Research, Endoscopic retrograde cholangiopancreatography -- Complications and side effects, Pancreatitis -- Prevention, Health
Published
2008

20. Estimating dissipation rates associated with double diffusion

Author

Middleton, L., Fine, E. C., MacKinnon, J. A., Alford, M. H., Taylor, J.R., Middleton, L., Fine, E. C., MacKinnon, J. A., Alford, M. H., and Taylor, J.R.

Abstract

Double diffusion refers to a variety of turbulent processes in which potential energy is released into kinetic energy, made possible in the ocean by the difference in molecular diffusivities between salinity and temperature. Here, we present a new method for estimating the kinetic energy dissipation rates forced by double-diffusive convection using temperature and salinity data alone. The method estimates the up-gradient diapycnal buoyancy flux associated with double diffusion, which is hypothesised to balance the dissipation rate. To calculate the temperature and salinity gradients on small scales we apply a canonical scaling for compensated thermohaline variance (or ‘spice’) on sub-measurement scales with a fixed buoyancy gradient. Our predicted dissipation rates compare favorably with microstructure measurements collected in the Chukchi Sea. Fine et al. (2018) showed that dissipation rates provide good estimates for heat fluxes in this region. Finally, we show the method maintains predictive skill when applied to a sub-sampling of the CTD data.

Published
2021

22. Optical modulation techniques for length sensing and control of optical cavities

Author

Barr, B.W., Huttner, S.H., Taylor, J.R., Sorazu, B., Plissi, M.V., and Strain, K.A.

Subjects
Amplitude modulation -- Methods, Holes -- Control, Length measurement -- Methods, Optical detectors -- Research, Astronomy, Physics
Abstract

We present a discussion of the use of amplitude modulation techniques with regard to the length sensing and control of optical cavities for laser interferometric gravitational-wave detectors. Traditional radio-frequency amplitude modulation techniques automatically include phase modulation as a product of the modulation process, which can contaminate the signal after demodulation. In particular, with many length-sensing and control schemes the detected signals are demodulated in quadrature, which, in the case of a traditional amplitude modulation scheme, will result in offsets due to the additional phase modulation. We demonstrate this effect using a simple optical cavity configuration and show that minor adjustments to the modulator system can be used to compensate for the extra modulation components and provide additional flexibility. OCIS codes: 230.0230, 230.2090, 230.4110, 230.5440.

Published
2007

23. Chromatin conformation and transcriptional activity are permissive regulators of DNA replication initiation in Drosophila

Author

David M. MacAlpine, Robin L. Armstrong, Taylor J.R. Penke, Robert J. Duronio, Daniel J. McKay, A. Gregory Matera, and Brian D. Strahl

Subjects
Male, Transcriptional Activation, 0301 basic medicine, X Chromosome, DNA replication initiation, Euchromatin, DNA Replication Timing, Heterochromatin, Biology, Histones, 03 medical and health sciences, Genetics, Animals, Genetics (clinical), Pericentric heterochromatin, Research, DNA replication, Chromatin Assembly and Disassembly, Chromosomes, Insect, Cell biology, Chromatin, 030104 developmental biology, Replication Initiation, Mutation, Drosophila, Female
Abstract

Chromatin structure has emerged as a key contributor to spatial and temporal control over the initiation of DNA replication. However, despite genome-wide correlations between early replication of gene-rich, accessible euchromatin and late replication of gene-poor, inaccessible heterochromatin, a causal relationship between chromatin structure and replication initiation remains elusive. Here, we combined histone gene engineering and whole-genome sequencing in Drosophila to determine how perturbing chromatin structure affects replication initiation. We found that most pericentric heterochromatin remains late replicating in H3K9R mutants, even though H3K9R pericentric heterochromatin is depleted of HP1a, more accessible, and transcriptionally active. These data indicate that HP1a loss, increased chromatin accessibility, and elevated transcription do not result in early replication of heterochromatin. Nevertheless, a small amount of pericentric heterochromatin with increased accessibility replicates earlier in H3K9R mutants. Transcription is de-repressed in these regions of advanced replication but not in those regions of the H3K9R mutant genome that replicate later, suggesting that transcriptional repression may contribute to late replication. We also explored relationships among chromatin, transcription, and replication in euchromatin by analyzing H4K16R mutants. In Drosophila, the X Chromosome gene expression is up-regulated twofold and replicates earlier in XY males than it does in XX females. We found that H4K16R mutation prevents normal male development and abrogates hyperexpression and earlier replication of the male X, consistent with previously established genome-wide correlations between transcription and early replication. In contrast, H4K16R females are viable and fertile, indicating that H4K16 modification is dispensable for genome replication and gene expression.

Published
2018
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24. Optophysiology: depth-resolved probing of retinal physiology with functional ultrahigh-resolution optical coherence tomography

Author

Bizheva, K., Pflug, R., Hermann, B., Povazay, B., Sattmann, H., Qiu, P., Anger, E., Reitsamer, H., Popov, S., Taylor, J.R., Unterhuber, A., Ahnelt, P., and Drexler, W.

Subjects
Photoreceptors -- Research, Science and technology
Abstract

Noncontact, depth-resolved, optical probing of retinal response to visual stimulation with a electroretinogram | functional optical coherence tomography | inner plexiform layer | photoreceptors | retinal imaging

Published
2006

29. Functional Redundancy of Variant and Canonical Histone H3 Lysine 9 Modification in Drosophila

Author

Robert J. Duronio, Daniel J. McKay, Taylor J.R. Penke, Brian D. Strahl, and A. Gregory Matera

Subjects
0301 basic medicine, Euchromatin, Heterochromatin, Biology, 03 medical and health sciences, Histone H3, 0302 clinical medicine, Histone H1, Histone H2A, Histone methylation, Transcriptional regulation, Genetics, Histone code, Epigenetics, 030304 developmental biology, Regulation of gene expression, 0303 health sciences, Chromatin, Cell biology, Histone, 030104 developmental biology, Histone methyltransferase, biology.protein, Heterochromatin protein 1, 030217 neurology & neurosurgery
Abstract

Histone post-translational modifications (PTMs) and differential incorporation of variant and canonical histones into chromatin are central modes of epigenetic regulation. Despite similar protein sequences, histone variants are enriched for different suites of PTMs compared to their canonical counterparts. For example, variant histone H3.3 occurs primarily in transcribed regions and is enriched for “active” histone PTMs like Lys9 acetylation (H3.3K9ac), whereas the canonical histone H3 is enriched for Lys9 methylation (H3K9me), which is found in transcriptionally silent heterochromatin. To determine the functions of K9 modification on variant versus canonical H3, we compared the phenotypes caused by engineering H3.3K9R and H3K9R mutant genotypes in Drosophila melanogaster. Whereas most H3.3K9R and a small number of H3K9R mutant animals are capable of completing development and do not have substantially altered protein coding transcriptomes, all H3.3K9RH3K9R combined mutants die soon after embryogenesis and display decreased expression of genes enriched for K9ac. These data suggest that the role of K9ac in gene activation during development can be provided by either H3 or H3.3. Conversely, we found that H3.3K9 is methylated at telomeric transposons, and this mark contributes to repressive chromatin architecture, supporting a role for H3.3 in heterochromatin that is distinct from that of H3. Thus, our genetic and molecular analyses demonstrate that K9 modification of variant and canonical H3 have overlapping roles in development and transcriptional regulation, though to differing extents in euchromatin and heterochromatin.

Published
2018
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38. Convectively driven mixed layer growth in a rotating, stratified fluid

Author

Ivey, G.N., Taylor, J.R., and Coates, M.J.

Subjects
Thermoclines (Oceanography) -- Research, Abyssal zone -- Research, Ocean-atmosphere interaction -- Research, Earth sciences
Abstract

For examining deep convective processes in a stratified ocean, a laboratory experiment generated a convective mixed layer by heating over the central portion of the base of a rotating, thermally stratified fluid and used bottom heating to to model cooling at the ocean surface. Results indicate that relatively unstratified deep water is ventilated in situations where the eddy does not become fully developed before the thermocline is penetrated.

Published
1995

39. Effects of vacuum exposure on stress and spectral shift of high reflective coatings

Author

Stolz, C.J., Taylor, J.R., Eickelberg, W.K., and Lindh, J.D.

Subjects
Optical coatings -- Research, Vacuum technology -- Analysis, Astronomy, Physics
Abstract

Coating stress and spectral shift are affected by changing from ambient to vacuum environments. This change can affect optical systems that are aligned in air and used in a vacuum or in a dry environment. Spectral shifts up to 3% and reflected wave-front changes up to 0.35 waves peak to valley are reported for conventional electron-beam deposition and ion-assisted deposition. Alternatively, ion-beam sputtered coatings have virtually no changes between different pressure environments.

Published
1993

42. H3K9 Promotes Under-Replication of Pericentromeric Heterochromatin in Drosophila Salivary Gland Polytene Chromosomes

Author

Robin L. Armstrong, Taylor J.R. Penke, Daniel J. McKay, Robert J. Duronio, Brian D. Strahl, Samuel K Chao, A. Gregory Matera, and Gabrielle M Gentile

Subjects
DNA Replication, 0301 basic medicine, lcsh:QH426-470, Euchromatin, Heterochromatin, Centromere, Methylation, Salivary Glands, Article, Endoreplication, Histones, 03 medical and health sciences, 0302 clinical medicine, Genetics, Animals, Endoreduplication, under-replication, Heterochromatin organization, Genetics (clinical), Pericentric heterochromatin, Polytene Chromosomes, Polytene chromosome, biology, fungi, H4K16, heterochromatin, Chromatin, Cell biology, lcsh:Genetics, H3K9, Drosophila melanogaster, 030104 developmental biology, Histone, biology.protein, Drosophila, Protein Processing, Post-Translational, 030217 neurology & neurosurgery
Abstract

Chromatin structure and its organization contributes to the proper regulation and timing of DNA replication. Yet, the precise mechanism by which chromatin contributes to DNA replication remains incompletely understood. This is particularly true for cell types that rely on polyploidization as a developmental strategy for growth and high biosynthetic capacity. During Drosophila larval development, cells of the salivary gland undergo endoreplication, repetitive rounds of DNA synthesis without intervening cell division, resulting in ploidy values of ~1350C. S phase of these endocycles displays a reproducible pattern of early and late replicating regions of the genome resulting from the activity of the same replication initiation factors that are used in diploid cells. However, unlike diploid cells, the latest replicating regions of polyploid salivary gland genomes, composed primarily of pericentric heterochromatic enriched in H3K9 methylation, are not replicated each endocycle, resulting in under-replicated domains with reduced ploidy. Here, we employ a histone gene replacement strategy in Drosophila to demonstrate that mutation of a histone residue important for heterochromatin organization and function (H3K9) but not mutation of a histone residue important for euchromatin function (H4K16), disrupts proper endoreplication in Drosophila salivary gland polyploid genomes thereby leading to DNA copy gain in pericentric heterochromatin. These findings reveal that H3K9 is necessary for normal levels of under-replication of pericentric heterochromatin and suggest that under-replication at pericentric heterochromatin is mediated through H3K9 methylation.

Published
2019
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47. An Animal Model for Genetic Analysis of Multi-Gene Families: Cloning and Transgenesis of Large Tandemly Repeated Histone Gene Clusters

Author

Robert J. Duronio, Taylor J.R. Penke, A. Gregory Matera, Mary Leatham-Jensen, Michael P. Meers, and Daniel J. McKay

Subjects
0301 basic medicine, biology, Transgene, Mutagenesis (molecular biology technique), Computational biology, biology.organism_classification, Genome, 03 medical and health sciences, 030104 developmental biology, Histone, biology.protein, Epigenetics, Drosophila melanogaster, Repeated sequence, Gene
Abstract

Histone post-translational modifications (PTMs) are thought to participate in a range of essential molecular and cellular processes, including gene expression, replication, and nuclear organization. Importantly, histone PTMs are also thought to be prime candidates for carriers of epigenetic information across cell cycles and generations. However, directly testing the necessity of histone PTMs themselves in these processes by mutagenesis has been extremely difficult to carry out because of the highly repetitive nature of histone genes in animal genomes. We developed a transgenic system to generate Drosophila melanogaster genotypes in which the entire complement of replication-dependent histone genes is mutant at a residue of interest. We built a BAC vector containing a visible marker for lineage tracking along with the capacity to clone large (60-100 kb) inserts that subsequently can be site-specifically integrated into the D. melanogaster genome. We demonstrate that artificial tandem arrays of the core 5 kb replication-dependent histone repeat can be generated with relative ease. This genetic platform represents the first histone replacement system to leverage a single tandem transgenic insertion for facile genetics and analysis of molecular and cellular phenotypes. We demonstrate the utility of our system for directly preventing histone residues from being modified, and studying the consequent phenotypes. This system can be generalized to the cloning and transgenic insertion of any tandemly repeated sequence of biological interest.

Published
2018
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