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Related Concept Videos

Transcription Factors02:16

Transcription Factors

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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Transcription01:10

Transcription

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Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
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Master Transcription Regulators02:23

Master Transcription Regulators

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Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
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Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

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Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
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Transcription Elongation Factors02:35

Transcription Elongation Factors

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Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
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Eukaryotic Transcription Inhibitors01:52

Eukaryotic Transcription Inhibitors

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Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells.
Eukaryotic transcription inhibitors usually contain two distinct domains, a...
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High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing
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eEF1A demonstrates paralog specific effects on HIV-1 reverse transcription efficiency.

Dongsheng Li1, Daniel J Rawle2, Zhonglan Wu3

  • 1Department of Cell and Molecular Biology, QIMR Berghofer Medical Research Institute, Herston, Qld 4006, Australia; Department of Genetics and Computational Biology, QIMR Berghofer Medical Research Institute, Herston, Qld 4006, Australia.

Virology
|February 21, 2019
PubMed
Summary

The eukaryotic translation elongation factor 1A (eEF1A) paralog eEF1A1, but not eEF1A2, supports HIV-1 reverse transcription. This suggests eEF1A1 is crucial for HIV-1 replication, unlike its paralog.

Keywords:
HEK 293T cellHIV-1Human primary skeletal muscle cellParalogReplicationReverses transcriptioneEF1A

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Area of Science:

  • Molecular Biology
  • Virology
  • Cell Biology

Background:

  • Eukaryotic translation elongation factor 1A (eEF1A) exists as two paralogs, eEF1A1 and eEF1A2, with shared roles in protein synthesis.
  • While eEF1A1 is implicated in viral replication, eEF1A2's role remains largely unexplored.
  • Previous work established eEF1A1's interaction with HIV-1 reverse transcriptase (RT) and its support for reverse transcription.

Purpose of the Study:

  • To investigate the differential roles of eEF1A1 and eEF1A2 in HIV-1 reverse transcription.
  • To compare the interaction affinities of eEF1A1 and eEF1A2 with HIV-1 RT.
  • To determine the correlation between eEF1A paralog expression levels and HIV-1 reverse transcription efficiency.

Main Methods:

  • Immunoprecipitation assays to assess protein-protein interactions.
  • Biolayer interferometry to quantify binding kinetics between eEF1A paralogs and HIV-1 RT.
  • Overexpression and knockdown experiments in HEK293T cells to evaluate functional roles.
  • Quantitative analysis of eEF1A mRNA levels in correlation with viral replication.

Main Results:

  • HIV-1 RT exhibited stronger interaction with eEF1A1 compared to eEF1A2.
  • Biolayer interferometry revealed distinct association and dissociation rates of eEF1A paralogs with RT.
  • Overexpression of eEF1A1, but not eEF1A2, restored HIV-1 reverse transcription efficiency in knockdown cells.
  • HIV-1 reverse transcription efficiency correlated positively with eEF1A1 mRNA levels, but not eEF1A2 mRNA levels, in both cell lines.

Conclusions:

  • eEF1A1 is specifically implicated in supporting HIV-1 reverse transcription, while eEF1A2 does not play a significant role.
  • The differential interaction and functional activity of eEF1A paralogs highlight distinct roles beyond canonical translation.
  • eEF1A1's expression level is a key determinant of HIV-1 reverse transcription efficiency, underscoring its importance in viral replication.