Mung bean nuclease I. 7. Terminally directed hydrolysis of native DNA

Kroeker, Warren D.; Kowalski, David; Laskowski, M.

doi:10.1021/bi00665a020

Cited by 74 publications

(31 citation statements)

References 20 publications

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“…Mung bean nuclease also digests single-stranded DNA or RNA. However, unlike S1 nuclease, it does not cleave the site opposite to the nick (22). Single-stranded overhangs of DNA can be a substrate for these enzymes.…”

Section: Discussionmentioning

confidence: 99%

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Enhancement of TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method using mung bean nuclease, a single-stranded DNA digestion enzyme.

Umemura

Yasuda²,

Osamura³

et al. 1996

J Histochem Cytochem.

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method has been employed widely to demonstrate apoptotic cells in routinely prepared paraffin sections. Because the apoptotic cells were reactive with the antibody to singlestranded DNA, we attempted to enhance the TUNEL positivity by pretreatment with single-stranded DNA digestion enzymes, S1 nuclease, and mung bean nuclease. When mung bean nuclease (5 Ul50 pllsection) was incubated at 37°C for 30 min, the TUNEL reaction was most effectively enhanced. Pretreatment with S1 nuclease (0.25 Ul50 pl/sec-

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Section: Discussionmentioning

confidence: 99%

“…An-other reason for the superiority of mung bean nuclease may relate to its specificity. Unlike S1 nuclease, mung bean nuclease cleaves overhanging bases immediately adjacent to the last hybridized pair without removing any of the base-paired nucleotides, to give precisely generated blunt ends (22).…”

Section: Discussionmentioning

confidence: 99%

Enhancement of TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method using mung bean nuclease, a single-stranded DNA digestion enzyme.

Umemura

Yasuda²,

Osamura³

et al. 1996

J Histochem Cytochem.

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“…As confirmed by sequencing, the DNA cloning site in pGBT9-2 is 5Ј-GGATTCCCGGGGATTCGACCTGCAGCC-3Ј. This plasmid was PvuII digested, treated with mung bean nuclease (29), and religated. In the resulting plasmid, pGBT9-3, the entire umuC coding sequence was cloned as described for pGBT9umuC.…”

Section: Methodsmentioning

confidence: 99%

Specific in vivo protein-protein interactions between Escherichia coli SOS mutagenesis proteins

Jonczyk

Nowicka

1996

J Bacteriol

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One of the components of the RecA-LexA-controlled SOS response in Escherichia coli cells is an inducible error-prone DNA replication pathway that results in a substantial increase in the mutation rate. It is believed that error-prone DNA synthesis is performed by a multiprotein complex that is formed by UmuC, UmuD, RecA, and probably DNA polymerase III holoenzyme. It is postulated that the formation of such a complex requires specific interactions between these proteins. We have analyzed the specific protein-protein interactions between UmuC, UmuD, and UmuD fusion proteins, using a Saccharomyces cerevisiae two-hybrid system. In agreement with previous in vitro data, we have shown that UmuD and UmuD are able to form both homodimers (UmuD-UmuD and UmuD-UmuD) and a heterodimer (UmuD-UmuD). Our data show that UmuC fusion protein is capable of interacting exclusively with UmuD and not with UmuD. Thus, posttranslational processing of UmuD into UmuD is a critical step in SOS mutagenesis, enabling only the latter protein to interact with UmuC. Our data seem to indicate that the integrity of the entire UmuC sequence is essential for UmuC-UmuD heterotypic interaction. Finally, in our studies, we used three different UmuC mutant proteins: UmuC25, UmuC36, and UmuC104. We have found that UmuC25 and UmuC36 are not capable of associating with UmuD. In contrast, UmuC104 protein interacts with UmuD protein with an efficiency identical to that of the wild-type protein. We postulate that UmuC104 protein might be defective in interaction with another, unknown protein essential for the SOS mutagenesis pathway.Exposure of Escherichia coli cells to agents that damage DNA and introduce replication-blocking lesions results in the induction of the SOS regulon (13,22,33,51,52). Approximately 20 cellular genes are derepressed when activated RecA promotes the proteolytic cleavage of LexA protein, the repressor of SOS genes (22,25,32,33,44,51,52).One of the components of the RecA-LexA-controlled SOS response is an inducible error-prone DNA replication pathway that causes a significant increase in the mutation rate (13,22,33,51,52). Genetic and physiological experiments indicate that the products of the umuDC operon and the recA gene are essential components of the SOS mutation pathway (3,12,16,17,26,44,47,48). This is supported by the fact that umuD and umuC mutants of E. coli are virtually nonmutable with UV light and many chemical agents (26,47).The recA and umuDC genes are members of the SOS regulon and are expressed at elevated levels after DNA damage (15,31,46,53,55). In SOS-competent cells, activated RecA also mediates the cleavage of UmuD at its Cys-24-Gly-25 bond, yielding a shorter UmuDЈ product, which has been proposed to be an active form of UmuD for mutagenesis (6,37,45,54). Several data suggest that UmuC, UmuDЈ, and RecA proteins facilitate the proceeding of the DNA replication complex beyond lesions in the template (5, 21, 40, 41). In addition, it has been shown that overexpression of the umuDC operon results in cold sensitivit...

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“…DNA-DNA hybridizations. Since some batches of S1 nuclease contain significant amounts of double-stranded DNase activity (14,16), the DNA hybrids were analyzed with mung bean nuclease [Pharmacia (Canada) Ltd.]. At low enzyme and high salt concentrations, mung bean nuclease will hydrolyze only single-stranded DNA (14,16).…”

Section: Methodsmentioning

confidence: 99%

Analysis of Actinobacillus pleuropneumoniae and Related Organisms by DNA-DNA Hybridization and Restriction Endonuclease Fingerprinting

Borr

Ryan

MacInnes

1991

International Journal of Systematic Bacteriology

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Mung bean nuclease I. 7. Terminally directed hydrolysis of native DNA

Cited by 74 publications

References 20 publications

Enhancement of TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method using mung bean nuclease, a single-stranded DNA digestion enzyme.

Enhancement of TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method using mung bean nuclease, a single-stranded DNA digestion enzyme.

Specific in vivo protein-protein interactions between Escherichia coli SOS mutagenesis proteins

Analysis of Actinobacillus pleuropneumoniae and Related Organisms by DNA-DNA Hybridization and Restriction Endonuclease Fingerprinting

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