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Related Concept Videos

Mechanism of Conjugation01:19

Mechanism of Conjugation

Bacterial conjugation is a mechanism of horizontal gene transfer that enables the exchange of genetic material between bacterial cells through direct contact. This process is facilitated by a donor cell carrying a conjugative plasmid, which encodes genes necessary for pilus formation, DNA replication, and transfer. The conjugative plasmid plays a central role in initiating and executing the transfer of genetic material.The tra region of the conjugative plasmid encodes proteins responsible for...
Conjugation01:19

Conjugation

Conjugation is a form of horizontal gene transfer that primarily occurs in bacteria and some archaea, promoting genetic diversity and adaptation. Bacteria can acquire resistance genes through conjugative plasmids, allowing them to survive antibiotic treatments that would otherwise be lethal. This process involves direct contact between cells through specialized structures such as the sex pilus and is mediated by conjugative plasmids, including the F (fertility) factor.Conjugation requires...
Phase II Conjugation Reactions: Overview01:14

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Conjugation, a key component of phase II biotransformation reactions, is a vital process in drug detoxification. It involves transferring endogenous substances like glucuronic acid, sulfate, and glycine to drugs or their metabolites formed in phase I reactions. These conjugation reactions, often catalyzed by specific enzymes, transform potentially harmful metabolites into inactive, water-soluble forms easily excreted in urine or bile. By enhancing polarity and eliminating pharmacological...
Phase II Reactions: Miscellaneous Conjugation Reactions01:19

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Phase II biotransformations are detoxification mechanisms that conjugate xenobiotics with endogenous substances, neutralizing their toxicity.
A key example involves the conjugation of cyanide ions, which impair cellular respiration and alter hemoglobin into non-oxygen-carrying cyanmethemoglobin. To neutralize this threat, a sulfur atom from thiosulphate is transferred to the cyanide ion, catalyzed by the enzyme rhodanese, resulting in an inactive compound called thiocyanate. The production of...
Enzyme-Linked Immunosorbent Assay01:33

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

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The ISG15 conjugation system.

Larissa A Durfee1, Jon M Huibregtse

  • 1Section of Molecular Genetics and Molecular Biology, Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 22, 2012
PubMed
Summary
This summary is machine-generated.

Interferon-stimulated gene 15 (ISG15) conjugation enzymes are expressed upon type 1 interferon signaling. Herc5 modifies proteins during translation, and this study details its expression and polysome association.

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Area of Science:

  • Molecular Biology
  • Immunology
  • Virology

Background:

  • Interferon-stimulated gene 15 (ISG15) is crucial for antiviral defense.
  • ISG15 conjugation is mediated by E1 (Ube1L), E2 (UbcH8), and E3 (Herc5) enzymes.
  • These enzymes are upregulated by type 1 interferon (IFN-α/β) signaling.

Purpose of the Study:

  • To investigate the expression of core ISG15 conjugating enzymes in human cells.
  • To describe methods for detecting ISG15 conjugates.
  • To analyze the association of Herc5 with polysomes for cotranslational modification.

Main Methods:

  • Analysis of core ISG15 conjugating enzyme expression.
  • Detection assays for ISG15 conjugates.
  • Cellular fractionation to isolate polysomes.
  • Co-immunoprecipitation to assess protein associations.

Main Results:

  • Core ISG15 conjugating enzymes (Ube1L, UbcH8, Herc5) are expressed in human cells.
  • IFN-α/β signaling induces the transcription of these enzymes.
  • Herc5 was found to associate with polysomes, indicating cotranslational modification of target proteins.

Conclusions:

  • The core ISG15ylation machinery is regulated by type 1 interferons.
  • Herc5 functions in a cotranslational manner, modifying proteins as they are synthesized.
  • This provides insight into the mechanisms of ISG15's role in antiviral responses.