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Related Experiment Videos

Multiple cis elements contribute to geminivirus origin function

B M Orozco1, H J Gladfelter, S B Settlage

  • 1Department of Biochemistry, North Carolina State University, Raleigh 27695-7622, USA. orozco@bchserver.bch.ncsu.edu

Virology
|March 26, 1998
PubMed
Summary
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Researchers studied the tomato golden mosaic virus (TGMV) replication origin. They found a minimal 89-bp region containing specific DNA sequences and structures is crucial for viral DNA replication.

Area of Science:

  • Molecular Biology
  • Virology
  • Plant Pathology

Background:

  • The tomato golden mosaic virus (TGMV) genome comprises two circular DNA molecules.
  • A conserved 200-bp common region is essential for TGMV replication.
  • Understanding the plus-strand origin is key to geminivirus replication mechanisms.

Purpose of the Study:

  • To characterize the minimal sequence and functional elements of the TGMV plus-strand origin.
  • To investigate the role of specific cis elements and viral replication proteins (AL1, AL3) in origin activity.
  • To elucidate the structural requirements for TGMV origin function in episomal replication.

Main Methods:

  • Analysis of TGMV common region sequences for episomal replication in tobacco protoplasts.
  • In trans supply of viral replication proteins AL1 and AL3.

Related Experiment Videos

  • Mutational analysis of origin sequences, including AL1 binding motifs and hairpin structures.
  • In vitro AL1/DNA binding assays.

    • Main Results:

    • A minimal 89-bp fragment containing an AL1 binding motif and a hairpin structure functions as the origin.
    • Two additional cis elements were identified: one essential for activity and another enhancing replication.
    • Both sequence and spacing of the AL1 binding site, as well as its distance from the hairpin, are critical for origin function.

    Conclusions:

    • The TGMV plus-strand origin is a complex regulatory region.
    • Multiple cis elements interact to facilitate the initiation of viral DNA replication.
    • Detailed understanding of TGMV replication can inform strategies against geminivirus infections.