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  1. Article ; Online: Long-range DNA end resection supports homologous recombination by checkpoint activation rather than extensive homology generation.

    Kimble, Michael T / Johnson, Matthew J / Nester, Mattie R / Symington, Lorraine S

    eLife

    2023  Volume 12

    Abstract: Homologous recombination (HR), the high-fidelity mechanism for double-strand break (DSB) repair, relies on DNA end resection by nucleolytic degradation of the 5'-terminated ends. However, the role of long-range resection mediated by Exo1 and/or Sgs1-Dna2 ...

    Abstract Homologous recombination (HR), the high-fidelity mechanism for double-strand break (DSB) repair, relies on DNA end resection by nucleolytic degradation of the 5'-terminated ends. However, the role of long-range resection mediated by Exo1 and/or Sgs1-Dna2 in HR is not fully understood. Here, we show that Exo1 and Sgs1 are dispensable for recombination between closely linked repeats, but are required for interchromosomal repeat recombination in
    MeSH term(s) Saccharomyces cerevisiae Proteins/genetics ; Saccharomyces cerevisiae Proteins/metabolism ; DNA Breaks, Double-Stranded ; DNA Repair ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Homologous Recombination ; Exodeoxyribonucleases/genetics ; Exodeoxyribonucleases/metabolism ; DNA/metabolism ; RecQ Helicases/metabolism
    Chemical Substances Saccharomyces cerevisiae Proteins ; Exodeoxyribonucleases (EC 3.1.-) ; DNA (9007-49-2) ; RecQ Helicases (EC 3.6.4.12)
    Language English
    Publishing date 2023-06-30
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2687154-3
    ISSN 2050-084X ; 2050-084X
    ISSN (online) 2050-084X
    ISSN 2050-084X
    DOI 10.7554/eLife.84322
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Genetic reporters to detect and quantify homologous recombination in yeast.

    Marie, Léa / Kimble, Michael T / Symington, Lorraine S

    Methods in cell biology

    2022  Volume 182, Page(s) 35–48

    Abstract: Homologous recombination is a conserved process that cells use to repair damaged DNA. Many genetic assays have been developed in Saccharomyces cerevisiae to measure and characterize different types of recombination events, as well as identify proteins ... ...

    Abstract Homologous recombination is a conserved process that cells use to repair damaged DNA. Many genetic assays have been developed in Saccharomyces cerevisiae to measure and characterize different types of recombination events, as well as identify proteins acting in such recombination events. Here, we describe two intrachromosomal reporters that utilize ade2 heteroalleles, whereby homologous recombination can be detected by colony color and adenine prototrophy. We detail the use of these reporters to measure recombination frequency, as well as to characterize the types of recombination events.
    MeSH term(s) Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Homologous Recombination/genetics ; DNA Damage ; Saccharomyces cerevisiae Proteins/genetics ; Saccharomyces cerevisiae Proteins/metabolism ; DNA Repair
    Chemical Substances Saccharomyces cerevisiae Proteins
    Language English
    Publishing date 2022-11-28
    Publishing country United States
    Document type Journal Article
    ISSN 0091-679X
    ISSN 0091-679X
    DOI 10.1016/bs.mcb.2022.10.011
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Sae2 antagonizes Rad9 accumulation at DNA double-strand breaks to attenuate checkpoint signaling and facilitate end resection.

    Yu, Tai-Yuan / Kimble, Michael T / Symington, Lorraine S

    Proceedings of the National Academy of Sciences of the United States of America

    2018  Volume 115, Issue 51, Page(s) E11961–E11969

    Abstract: The Mre11-Rad50- ... ...

    Abstract The Mre11-Rad50-Xrs2
    MeSH term(s) Cell Cycle/genetics ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Checkpoint Kinase 2/metabolism ; DNA/metabolism ; DNA Breaks, Double-Stranded ; DNA Damage ; DNA Helicases ; DNA Repair/physiology ; DNA, Fungal/genetics ; DNA, Fungal/metabolism ; DNA-Binding Proteins/metabolism ; Endodeoxyribonucleases/genetics ; Endodeoxyribonucleases/metabolism ; Endonucleases/genetics ; Endonucleases/metabolism ; Exodeoxyribonucleases/genetics ; Exodeoxyribonucleases/metabolism ; Intracellular Signaling Peptides and Proteins/metabolism ; Mutagens ; Protein-Serine-Threonine Kinases/metabolism ; RecQ Helicases/genetics ; RecQ Helicases/metabolism ; Saccharomyces cerevisiae/metabolism ; Saccharomyces cerevisiae Proteins/genetics ; Saccharomyces cerevisiae Proteins/metabolism ; Signal Transduction/genetics
    Chemical Substances Cell Cycle Proteins ; DNA, Fungal ; DNA-Binding Proteins ; Intracellular Signaling Peptides and Proteins ; Mutagens ; RAD50 protein, S cerevisiae ; SAE2 protein, S cerevisiae ; Saccharomyces cerevisiae Proteins ; rad9 protein (139691-42-2) ; DNA (9007-49-2) ; Checkpoint Kinase 2 (EC 2.7.1.11) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; TEL1 protein, S cerevisiae (EC 2.7.11.1) ; RAD53 protein, S cerevisiae (EC 2.7.12.1) ; Endodeoxyribonucleases (EC 3.1.-) ; Endonucleases (EC 3.1.-) ; Exodeoxyribonucleases (EC 3.1.-) ; MRE11 protein, S cerevisiae (EC 3.1.-) ; exodeoxyribonuclease I (EC 3.1.11.1) ; SGS1 protein, S cerevisiae (EC 3.6.1.-) ; DNA Helicases (EC 3.6.4.-) ; DNA2 protein, S cerevisiae (EC 3.6.4.12) ; RecQ Helicases (EC 3.6.4.12)
    Language English
    Publishing date 2018-12-03
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.1816539115
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  4. Article ; Online: An in vivo method for diversifying the functions of therapeutic antibodies.

    Tian, Ming / Cheng, Hwei-Ling / Kimble, Michael T / McGovern, Kelly / Waddicor, Peyton / Chen, Yiwei / Cantor, Elizabeth / Qiu, Mengting / Tuchel, Marie-Elen / Dao, Mai / Alt, Frederick W

    Proceedings of the National Academy of Sciences of the United States of America

    2021  Volume 118, Issue 10

    Abstract: V(D)J recombination generates mature B cells that express huge repertoires of primary antibodies as diverse immunoglobulin (Ig) heavy chain (IgH) and light chain (IgL) of their B cell antigen receptors (BCRs). Cognate antigen binding to BCR variable ... ...

    Abstract V(D)J recombination generates mature B cells that express huge repertoires of primary antibodies as diverse immunoglobulin (Ig) heavy chain (IgH) and light chain (IgL) of their B cell antigen receptors (BCRs). Cognate antigen binding to BCR variable region domains activates B cells into the germinal center (GC) reaction in which somatic hypermutation (SHM) modifies primary variable region-encoding sequences, with subsequent selection for mutations that improve antigen-binding affinity, ultimately leading to antibody affinity maturation. Based on these principles, we developed a humanized mouse model approach to diversify an anti-PD1 therapeutic antibody and allow isolation of variants with novel properties. In this approach, component Ig gene segments of the anti-PD1 antibody underwent de novo V(D)J recombination to diversify the anti-PD1 antibody in the primary antibody repertoire in the mouse models. Immunization of these mouse models further modified the anti-PD1 antibodies through SHM. Known anti-PD1 antibodies block interaction of PD1 with its ligands to alleviate PD1-mediated T cell suppression, thereby boosting antitumor T cell responses. By diversifying one such anti-PD1 antibody, we derived many anti-PD1 antibodies, including anti-PD1 antibodies with the opposite activity of enhancing PD1/ligand interaction. Such antibodies theoretically might suppress deleterious T cell activities in autoimmune diseases. The approach we describe should be generally applicable for diversifying other therapeutic antibodies.
    MeSH term(s) Animals ; Antibody Affinity/genetics ; Immunoglobulin Heavy Chains/genetics ; Immunoglobulin Heavy Chains/immunology ; Immunoglobulin Light Chains/genetics ; Immunoglobulin Light Chains/immunology ; Mice ; Receptors, Antigen, B-Cell/genetics ; Receptors, Antigen, B-Cell/immunology ; Somatic Hypermutation, Immunoglobulin ; V(D)J Recombination/immunology
    Chemical Substances Immunoglobulin Heavy Chains ; Immunoglobulin Light Chains ; Receptors, Antigen, B-Cell
    Language English
    Publishing date 2021-06-11
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.2025596118
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  5. Article ; Online: Conditional antibody expression to avoid central B cell deletion in humanized HIV-1 vaccine mouse models.

    Tian, Ming / McGovern, Kelly / Cheng, Hwei-Ling / Waddicor, Peyton / Rieble, Lisa / Dao, Mai / Chen, Yiwei / Kimble, Michael T / Cantor, Elizabeth / Manfredonia, Nicole / Judson, Rachael / Chapdelaine-Williams, Aimee / Cain, Derek W / Haynes, Barton F / Alt, Frederick W

    Proceedings of the National Academy of Sciences of the United States of America

    2020  Volume 117, Issue 14, Page(s) 7929–7940

    Abstract: HIV-1 vaccine development aims to elicit broadly neutralizing antibodies (bnAbs) against diverse viral strains. In some HIV-1-infected individuals, bnAbs evolved from precursor antibodies through affinity maturation. To induce bnAbs, a vaccine must ... ...

    Abstract HIV-1 vaccine development aims to elicit broadly neutralizing antibodies (bnAbs) against diverse viral strains. In some HIV-1-infected individuals, bnAbs evolved from precursor antibodies through affinity maturation. To induce bnAbs, a vaccine must mediate a similar antibody maturation process. One way to test a vaccine is to immunize mouse models that express human bnAb precursors and assess whether the vaccine can convert precursor antibodies into bnAbs. A major problem with such mouse models is that bnAb expression often hinders B cell development. Such developmental blocks may be attributed to the unusual properties of bnAb variable regions, such as poly-reactivity and long antigen-binding loops, which are usually under negative selection during primary B cell development. To address this problem, we devised a method to circumvent such B cell developmental blocks by expressing bnAbs conditionally in mature B cells. We validated this method by expressing the unmutated common ancestor (UCA) of the human VRC26 bnAb in transgenic mice. Constitutive expression of the VRC26UCA led to developmental arrest of B cell progenitors in bone marrow; poly-reactivity of the VRC26UCA and poor pairing of the VRC26UCA heavy chain with the mouse surrogate light chain may contribute to this phenotype. The conditional expression strategy bypassed the impediment to VRC26UCA B cell development, enabling the expression of VRC26UCA in mature B cells. This approach should be generally applicable for expressing other bnAbs that are under negative selection during B cell development.
    MeSH term(s) AIDS Vaccines/immunology ; AIDS Vaccines/pharmacology ; Animals ; Antibodies, Neutralizing/immunology ; Antibodies, Neutralizing/physiology ; B-Lymphocytes/immunology ; B-Lymphocytes/virology ; Disease Models, Animal ; HIV Antibodies/immunology ; HIV Antibodies/pharmacology ; HIV Infections/immunology ; HIV Infections/prevention & control ; HIV Infections/virology ; HIV Seropositivity/genetics ; HIV Seropositivity/immunology ; HIV-1/drug effects ; HIV-1/immunology ; HIV-1/pathogenicity ; Humans ; Lymphocyte Activation/immunology ; Mice ; env Gene Products, Human Immunodeficiency Virus/genetics ; env Gene Products, Human Immunodeficiency Virus/immunology
    Chemical Substances AIDS Vaccines ; Antibodies, Neutralizing ; HIV Antibodies ; env Gene Products, Human Immunodeficiency Virus
    Language English
    Publishing date 2020-03-24
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.1921996117
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

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