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Related Concept Videos

LTR Retrotransposons03:08

LTR Retrotransposons

LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
Retrovirus Life Cycles01:10

Retrovirus Life Cycles

Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the retrovirus to...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...

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Related Experiment Video

Updated: Jul 13, 2026

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
13:07

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells

Published on: January 30, 2019

Ty1 reverse transcriptase does not read through the proposed 2',5'-branched retrotransposition intermediate in vitro.

Elizabeth D Pratico1, Scott K Silverman

  • 1Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

RNA (New York, N.Y.)
|July 27, 2007
PubMed
Summary

Ty1 retrotransposition does not require branched RNA intermediates. Ty1 reverse transcriptase poorly reads through proposed branch sites, challenging the obligatory role of branched RNA in Ty1 cDNA synthesis.

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Last Updated: Jul 13, 2026

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
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Detection of Retrotransposition Activity of Hot LINE-1s by Long-Distance Inverse PCR
10:54

Detection of Retrotransposition Activity of Hot LINE-1s by Long-Distance Inverse PCR

Published on: July 27, 2019

Area of Science:

  • Molecular Biology
  • Retrotransposition Mechanisms
  • Enzymology

Background:

  • 2',5'-branched RNA was proposed as a crucial intermediate in Ty1 retrotransposition.
  • Cleavage of branched RNA by lariat debranching enzyme (Dbr1p) was thought to enable complete Ty1 cDNA synthesis.

Purpose of the Study:

  • To directly test the ability of Ty1 reverse transcriptase (RT) to read through the proposed 2',5'-branched RNA branch site.
  • To evaluate the efficiency of Ty1 RT read-through in an in vitro system.

Main Methods:

  • Synthesized 2',5'-branched RNA corresponding to the proposed Ty1 branch site using deoxyribozymes.
  • Utilized an in vitro assay with purified Ty1 RT, TyA chaperone, and other necessary components.
  • Assessed strand transfer and read-through efficiency with RNase H+ and RNase H- Ty1 RT variants.

Main Results:

  • Ty1 RT elongates DNA synthesis up to the branch site.
  • Strand transfer occurs with RNase H+ Ty1 RT but not with RNase H- variant.
  • Ty1 RT exhibits extremely low read-through efficiency (<0.3%) at the branch site.

Conclusions:

  • The low read-through efficiency contradicts the hypothesis of branched RNA as an obligatory Ty1 intermediate.
  • Dbr1p's role in Ty1 retrotransposition may not involve 2',5'-phosphodiesterase activity.
  • Alternative mechanisms for Ty1 cDNA synthesis in dbr1 cells need to be explored.