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Ro60 ribonucleoprotein inhibits transcription by T3 RNA polymerase in vitro
M D Campos-Almaraz1, O Barbosa-Cisneros, R Herrera-Esparza
1Department of Molecular Biology and Immunotechnology, Universidad Autonoma de Zacatecas, Guadalupe, Mexico.
Background:
Ro60 ribonucleoprotein is a conserved molecule belonging to the family of Ro ribonucleoproteins and targeted by autoantibodies produced in patients with systemic lupus erythematosus or Sjogren's syndrome. Ro60 plays a role in postranscription events, as well as in the nucleocytoplasmic shuttling of RNA polymerase III transcripts.
Objective:
To evaluate the in vitro effects of Ro60 on T3 RNA polymerase transcription.
Methods:
Ro60 ribonucleoprotein was affinity-purified from human spleen extracts using 4B-Sepharose linked to anti-Ro60 monoclonal antibodies. Purified Ro60 was incorporated into the T3 RNA polymerase transcription reaction system using pTRI-beta-Actin-human DNA as a template.
Results:
Ro60 inhibited initiation of the transcription process in a dose-dependent manner; neither elongation nor termination was affected by Ro.
Conclusion:
In vitro, Ro60 appears to inhibit the transcription of a T3 RNA polymerase-dependent template.