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https://www.readbyqxmd.com/read/24236613/virulence-and-transferability-of-resistance-determinants-in-a-novel-klebsiella-pneumoniae-sequence-type-1137-in-china
#1
Yang Liu, Xiang-Yang Li, La-Gen Wan, Wei-Yan Jiang, Jing-Hong Yang, Fang-Qu Li
A study was designed to characterize three carbapenemase-producing Klebsiella pneumoniae isolated from pediatric patients in China. Molecular characterization was done using polymerase chain reaction and sequencing for blaVIM, blaNDM, blaIMP, blaKPC, blaCTX-Ms, blaOXAs, blaTEMs, and blaSHV; plasmid-mediated quinolone resistance determinants; aminoglycoside resistance determinants; multilocus sequencing typing; plasmid replicon typing; addiction; and virulence factors. Kp32 belonged to the newly described sequence type 1137, were positive for aac(6')-Ib-suzhou, qnrA1, qnrB4, qnrS1, aac(6')-Ib, rmtB, armA, blaSHV-12, blaCTX-M-15, blaKPC-2, and blaIMP-4; contained IncA/C plasmids that tested positive for K1 capsular antigens, the ccdAB (coupled cell division locus) addiction system and the wabG, ureA, rmpA, magA, allS, fimH, and the aerobactin virulence factors...
April 2014: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
https://www.readbyqxmd.com/read/24191025/dna-polymerase-%C3%AE-interacting-protein-2-is-a-processivity-factor-for-dna-polymerase-%C3%AE-during-8-oxo-7-8-dihydroguanine-bypass
#2
Giovanni Maga, Emmanuele Crespan, Enni Markkanen, Ralph Imhof, Antonia Furrer, Giuseppe Villani, Ulrich Hübscher, Barbara van Loon
The bypass of DNA lesions by the replication fork requires a switch between the replicative DNA polymerase (Pol) and a more specialized translesion synthesis (TLS) Pol to overcome the obstacle. DNA Pol δ-interacting protein 2 (PolDIP2) has been found to physically interact with Pol η, Pol ζ, and Rev1, suggesting a possible role of PolDIP2 in the TLS reaction. However, the consequences of PolDIP2 interaction on the properties of TLS Pols remain unknown. Here, we analyzed the effects of PolDIP2 on normal and TLS by five different human specialized Pols from three families: Pol δ (family B), Pol η and Pol ι (family Y), and Pol λ and Pol β (family X)...
November 19, 2013: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/23118481/silencing-of-human-dna-polymerase-%C3%AE-causes-replication-stress-and-is-synthetically-lethal-with-an-impaired-s-phase-checkpoint
#3
Elisa Zucca, Federica Bertoletti, Ursula Wimmer, Elena Ferrari, Giuliano Mazzini, Svetlana Khoronenkova, Nicole Grosse, Barbara van Loon, Grigory Dianov, Ulrich Hübscher, Giovanni Maga
Human DNA polymerase (pol) λ functions in base excision repair and non-homologous end joining. We have previously shown that DNA pol λ is involved in accurate bypass of the two frequent oxidative lesions, 7,8-dihydro-8-oxoguanine and 1,2-dihydro-2-oxoadenine during the S phase. However, nothing is known so far about the relationship of DNA pol λ with the S phase DNA damage response checkpoint. Here, we show that a knockdown of DNA pol λ, but not of its close homologue DNA pol β, results in replication fork stress and activates the S phase checkpoint, slowing S phase progression in different human cancer cell lines...
January 7, 2013: Nucleic Acids Research
https://www.readbyqxmd.com/read/23101935/human-dna-polymerase-%C3%AE-but-not-%C3%AE-can-bypass-a-2-deoxyribonolactone-lesion-together-with-proliferating-cell-nuclear-antigen
#4
Emmanuele Crespan, Emanuela Pasi, Shuhei Imoto, Ulrich Hübscher, Marc M Greenberg, Giovanni Maga
The C1'-oxidized lesion 2-deoxyribonolactone (L) is induced by free radical attack of DNA. This lesion is mutagenic, inhibits base excision repair, and can lead to strand scission. In double-stranded DNA L is repaired by long-patch base excision repair, but it induces replication fork arrest in a single-strand template. Translesion synthesis requires a specialized DNA polymerase (Pol). In E. coli, Pol V is responsible for bypassing L, whereas in yeast Pol ζ has been shown to be required for efficient bypass...
February 15, 2013: ACS Chemical Biology
https://www.readbyqxmd.com/read/22373917/microhomology-mediated-dna-strand-annealing-and-elongation-by-human-dna-polymerases-%C3%AE-and-%C3%AE-on-normal-and-repetitive-dna-sequences
#5
Emmanuele Crespan, Tibor Czabany, Giovanni Maga, Ulrich Hübscher
'Classical' non-homologous end joining (NHEJ), dependent on the Ku70/80 and the DNA ligase IV/XRCC4 complexes, is essential for the repair of DNA double-strand breaks. Eukaryotic cells possess also an alternative microhomology-mediated end-joining (MMEJ) mechanism, which is independent from Ku and DNA ligase 4/XRCC4. The components of the MMEJ machinery are still largely unknown. Family X DNA polymerases (pols) are involved in the classical NHEJ pathway. We have compared in this work, the ability of human family X DNA pols β, λ and μ, to promote the MMEJ of different model templates with terminal microhomology regions...
July 2012: Nucleic Acids Research
https://www.readbyqxmd.com/read/21325140/oxidative-dna-damage-bypass-in-arabidopsis-thaliana-requires-dna-polymerase-%C3%AE-and-proliferating-cell-nuclear-antigen-2
#6
Alessandra Amoroso, Lorenzo Concia, Caterina Maggio, Cécile Raynaud, Catherine Bergounioux, Emmanuele Crespan, Rino Cella, Giovanni Maga
The oxidized base 7,8-oxoguanine (8-oxo-G) is the most common DNA lesion generated by reactive oxygen species. This lesion is highly mutagenic due to the frequent misincorporation of A opposite 8-oxo-G during DNA replication. In mammalian cells, the DNA polymerase (pol) family X enzyme DNA pol λ catalyzes the correct incorporation of C opposite 8-oxo-G, together with the auxiliary factor proliferating cell nuclear antigen (PCNA). Here, we show that Arabidopsis thaliana DNA pol λ, the only member of the X family in plants, is as efficient in performing error-free translesion synthesis past 8-oxo-G as its mammalian homolog...
February 2011: Plant Cell
https://www.readbyqxmd.com/read/20423048/dna-polymerases-beta-and-lambda-bypass-thymine-glycol-in-gapped-dna-structures
#7
COMPARATIVE STUDY
Ekaterina A Belousova, Giovanni Maga, Yang Fan, Elena A Kubareva, Elena A Romanova, Natalia A Lebedeva, Tatiana S Oretskaya, Olga I Lavrik
Here we investigated the ability of the human X-family DNA polymerases beta and lambda to bypass thymine glycol (Tg) in gapped DNA substrates with the damage located in a defined position of the template strand. Maximum velocities and the Michaelis constant values were determined to study DNA synthesis in the presence of either Mg(2+) or Mn(2+). Additionally, the influence of hRPA (human replication protein A) and hPCNA (human proliferating cell nuclear antigen) on TLS (translesion synthesis) activity of DNA polymerases beta and lambda was examined...
June 8, 2010: Biochemistry
https://www.readbyqxmd.com/read/19329428/the-block-of-dna-polymerase-delta-strand-displacement-activity-by-an-abasic-site-can-be-rescued-by-the-concerted-action-of-dna-polymerase-beta-and-flap-endonuclease-1
#8
Giovanni Maga, Barbara van Loon, Emmanuele Crespan, Giuseppe Villani, Ulrich Hübscher
Abasic (AP) sites are very frequent and dangerous DNA lesions. Their ability to block the advancement of a replication fork has been always viewed as a consequence of their inhibitory effect on the DNA synthetic activity of replicative DNA polymerases (DNA pols). Here we show that AP sites can also affect the strand displacement activity of the lagging strand DNA pol delta, thus preventing proper Okazaki fragment maturation. This block can be overcome through a polymerase switch, involving the combined physical and functional interaction of DNA pol beta and Flap endonuclease 1...
May 22, 2009: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/19201975/reciprocal-expression-of-integration-host-factor-and-hu-in-the-developmental-cycle-and-infectivity-of-legionella-pneumophila
#9
Michael G Morash, Ann Karen C Brassinga, Michelle Warthan, Poornima Gourabathini, Rafael A Garduño, Steven D Goodman, Paul S Hoffman
Legionella pneumophila is an intracellular parasite of protozoa that differentiates late in infection into metabolically dormant cysts that are highly infectious. Regulation of this process is poorly understood. Here we report that the small DNA binding regulatory proteins integration host factor (IHF) and HU are reciprocally expressed over the developmental cycle, with HU expressed during exponential phase and IHF expressed postexponentially. To assess the role of these regulatory proteins in development, chromosomal deletions were constructed...
April 2009: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/19104052/replication-protein-a-and-proliferating-cell-nuclear-antigen-coordinate-dna-polymerase-selection-in-8-oxo-guanine-repair
#10
Giovanni Maga, Emmanuele Crespan, Ursula Wimmer, Barbara van Loon, Alessandra Amoroso, Chiara Mondello, Cristina Belgiovine, Elena Ferrari, Giada Locatelli, Giuseppe Villani, Ulrich Hübscher
The adenine misincorporated by replicative DNA polymerases (pols) opposite 7,8-dihydro-8-oxoguanine (8-oxo-G) is removed by a specific glycosylase, leaving the lesion on the DNA. Subsequent incorporation of C opposite 8-oxo-G on the resulting 1-nt gapped DNA is essential for the removal of the 8-oxo-G to prevent G-C to T-A transversion mutations. By using model DNA templates, purified DNA pols beta and lambda and knockout cell extracts, we show here that the auxiliary proteins replication protein A and proliferating cell nuclear antigen act as molecular switches to activate the DNA pol lambda- dependent highly efficient and faithful repair of A:8-oxo-G mismatches in human cells and to repress DNA pol beta activity...
December 30, 2008: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/17666409/error-free-bypass-of-2-hydroxyadenine-by-human-dna-polymerase-lambda-with-proliferating-cell-nuclear-antigen-and-replication-protein-a-in-different-sequence-contexts
#11
Emmanuele Crespan, Ulrich Hübscher, Giovanni Maga
1,2-dihydro-2-oxoadenine (2-OH-A), a common DNA lesion produced by reactive oxygen species, is a strong replicative block for several DNA polymerases (DNA pols). We have previously shown that various bases can be misincorporated opposite the 2-OH-A lesion and the type of mispairs varies with either the sequence context or the type of DNA pol tested. Here, we have analysed the ability of the human pol family X member DNA pol lambda, to bypass the 2-OH-A lesion. DNA pol lambda can perform error-free bypass of 2-OH-A when this lesion is located in a random sequence, whereas in a repeated sequence context, even though bypass was also largely error-free, misincorporation of dGMP could be observed...
2007: Nucleic Acids Research
https://www.readbyqxmd.com/read/17507928/8-oxo-guanine-bypass-by-human-dna-polymerases-in-the-presence-of-auxiliary-proteins
#12
Giovanni Maga, Giuseppe Villani, Emmanuele Crespan, Ursula Wimmer, Elena Ferrari, Barbara Bertocci, Ulrich Hübscher
Specialized DNA polymerases (DNA pols) are required for lesion bypass in human cells. Auxiliary factors have an important, but so far poorly understood, role. Here we analyse the effects of human proliferating cell nuclear antigen (PCNA) and replication protein A (RP-A) on six different human DNA pols--belonging to the B, Y and X classes--during in vitro bypass of different lesions. The mutagenic lesion 8-oxo-guanine (8-oxo-G) has high miscoding potential. A major and specific effect was found for 8-oxo-G bypass with DNA pols lambda and eta...
May 31, 2007: Nature
https://www.readbyqxmd.com/read/17188944/replication-of-2-hydroxyadenine-containing-dna-and-recognition-by-human-mutsalpha
#13
Flavia Barone, Scott D McCulloch, Peter Macpherson, Giovanni Maga, Masami Yamada, Takehiko Nohmi, Anna Minoprio, Filomena Mazzei, Thomas A Kunkel, Peter Karran, Margherita Bignami
2-Hydroxyadenine (2-OH-A), a product of DNA oxidation, is a potential source of mutations. We investigated how representative DNA polymerases from the A, B and Y families dealt with 2-OH-A in primer extension experiments. A template 2-OH-A reduced the rate of incorporation by DNA polymerase alpha (Pol alpha) and Klenow fragment (Kf(exo-)). Two Y family DNA polymerases, human polymerase eta (Pol eta) and the archeal Dpo4 polymerase were affected differently. Bypass by Pol eta was very inefficient whereas Dpo4 efficiently replicated 2-OH-A...
March 1, 2007: DNA Repair
https://www.readbyqxmd.com/read/16918360/human-terminal-deoxynucleotidyl-transferases-as-novel-targets-for-anticancer-chemotherapy
#14
REVIEW
Roberto Di Santo, Giovanni Maga
Mammalian terminal deoxyribonucleotidyl transferase (TDT) catalyzes the non-template-directed polymerization of deoxyribonucleoside triphosphates and has a key role in V(D)J recombination during lymphocyte and repertoire development. Over 90% of leukemic cells in acute lymphocytic leukemia and approximately 30% of leukemic cells in the chronic myelogenous leukemia crisis show elevated TDT activity. This finding is connected to a poor prognosis and response to chemotherapy and reduced survival time. On the other hand, recent data indicated that TDT is not the only terminal deoxyribonucleotidyl transferase in mammalian cells...
2006: Current Medicinal Chemistry
https://www.readbyqxmd.com/read/16522650/human-replication-protein-a-can-suppress-the-intrinsic-in-vitro-mutator-phenotype-of-human-dna-polymerase-lambda
#15
Giovanni Maga, Igor Shevelev, Giuseppe Villani, Silvio Spadari, Ulrich Hübscher
DNA polymerase lambda (pol lambda) is a member of the X family DNA polymerases and is endowed with multiple enzymatic activities. In this work we investigated the in vitro miscoding properties of full-length, human pol lambda either in the absence or in the presence of the human auxiliary proteins proliferating cell nuclear antigen (PCNA) and replication protein A (RP-A). Our data suggested that (i) pol lambda had an intrinsic ability to create mismatches and to incorporate ribonucleotides at nearly physiological Mn++ and Mg++ concentrations; (ii) the sequence of the template-primer could influence the misincorporation frequency of pol lambda; (iii) pol lambda preferentially generated G:T and G:G mismatches; (iv) RP-A, but not PCNA, selectively prevented misincorporation of an incorrect nucleotide by pol lambda, without affecting correct incorporation and (v) this inhibitory effect required a precise ratio between the concentrations of pol lambda and RP-A...
2006: Nucleic Acids Research
https://www.readbyqxmd.com/read/15992031/molecular-basis-for-the-antiviral-and-anticancer-activities-of-unnatural-l-beta-nucleosides
#16
S Spadari, G Maga, A Verri, F Focher
As a general rule, enzymes act on only one enantiomer of a chiral substrate and only one of the enantiomeric forms of a chiral molecule may bind effectively at the catalytic site, displaying biological activity. In recent years, some exceptions have been found among viral and cellular enzymes involved in the synthesis of deoxynucleoside triphosphates and in their polymerisation into DNA. Examples are: herpes virus thymidine kinases, cellular deoxycytidine kinase and deoxynucleotide kinases, human immunodeficiency virus type 1 (HIV-1) reverse transcriptase, hepatitis B virus (HBV) DNA polymerase and, to a lesser extent, some cellular DNA polymerases...
August 1998: Expert Opinion on Investigational Drugs
https://www.readbyqxmd.com/read/15729726/effect-of-differences-in-cancer-cells-and-tumor-growth-sites-on-recruiting-bone-marrow-derived-endothelial-cells-and-myofibroblasts-in-cancer-induced-stroma
#17
Takafumi Sangai, Genichiro Ishii, Keiji Kodama, Shin'ichi Miyamoto, Yasuyuki Aoyagi, Takashi Ito, Junji Magae, Hiroki Sasaki, Takeshi Nagashima, Masaru Miyazaki, Atsushi Ochiai
Cancer-stromal interaction is well known to play important roles during cancer progression. Recently we have demonstrated that bone marrow-derived vascular endothelial cells (BMD-VE) and myofibroblasts (BMD-MF) are recruited into the human pancreatic cancer cell line Capan-1 induced stroma. To assess the effect of the difference in cancer cell types on the recruitment of BMD-VE and BMD-MF, 10 kinds of human cancer cell line were implanted into the subcutaneous tissue of the immunodeficient mice transplanted with bone marrow of double-mutant mice (RAG-1-/- beta-gal Tg or RAG-1-/- GFP Tg)...
July 20, 2005: International Journal of Cancer. Journal International du Cancer
https://www.readbyqxmd.com/read/15537631/dna-elongation-by-the-human-dna-polymerase-lambda-polymerase-and-terminal-transferase-activities-are-differentially-coordinated-by-proliferating-cell-nuclear-antigen-and-replication-protein-a
#18
Giovanni Maga, Kristijan Ramadan, Giada A Locatelli, Igor Shevelev, Silvio Spadari, Ulrich Hübscher
DNA polymerase lambda contains template-dependent (DNA polymerase) and template-independent (terminal transferase) activities. In this study we enzymologically characterized the terminal transferase activity of polymerase lambda (pol lambda-tdt). Pol lambda-tdt activity was strongly influenced by the nature of the 3'-terminal sequence of the DNA substrate, and it required a single-stranded (ss) DNA 3'-overhang of about 9-12 nucleotides for optimal activity. The strong preference observed for pyrimidine versus purine nucleotide incorporation was found to be due, at least partially, to a steric block imposed by the residue Tyr-505 in the active site of pol lambda...
January 21, 2005: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/15377629/hot-topic-using-a-stearoyl-coa-desaturase-transgene-to-alter-milk-fatty-acid-composition
#19
W A Reh, E A Maga, N M B Collette, A Moyer, J S Conrad-Brink, S J Taylor, E J DePeters, S Oppenheim, J D Rowe, R H BonDurant, G B Anderson, J D Murray
Stearoyl-CoA desaturase enzyme converts specific medium- and long-chain saturated fatty acids to their monounsaturated form. Transgenic goats expressing a bovine beta-lactoglobulin promoter-rat stearoyl-CoA desaturase cDNA construct in mammary gland epithelial cells were produced by pronuclear microinjection. The fatty acid composition of milk from 4 female transgenic founders was analyzed on d 7, 14, and 30 of their first lactation. In 2 animals, the expression of the transgene changed the overall fatty acid composition of the resulting milk fat to a less saturated and more monounsaturated fatty acid profile at d 7 of lactation; however, this effect diminished by d 30...
October 2004: Journal of Dairy Science
https://www.readbyqxmd.com/read/15358682/the-human-dna-polymerase-lambda-interacts-with-pcna-through-a-domain-important-for-dna-primer-binding-and-the-interaction-is-inhibited-by-p21-waf1-cip1
#20
Giovanni Maga, Giuseppina Blanca, Igor Shevelev, Isabelle Frouin, Kristijan Ramadan, Silvio Spadari, Giuseppe Villani, Ulrich Hübscher
In this paper we show that DNA polymerase lambda (pol lambda) interacts with proliferating cell nuclear antigen (PCNA) in vivo in human cells. Moreover, by using recombinant mutated PCNA, we could demonstrate that pol lambda interacts with both the interdomain-connecting loop and the nearby hydrophobic pocket on the anterior of PCNA and that critical residues within a helix-hairpin-helix domain of pol lambda, important for proper DNA primer binding, are also involved in the enzyme's interaction with PCNA. Finally, we show that the tumor suppressor protein p21(WAF1/CIP1) can efficiently compete in vitro with pol lambda for binding to PCNA...
November 2004: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
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