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Gene by Promoter-Proximal Pausing of RNA Polymerase II1
Institute for Clinical Molecular Biology and Tumor Genetics, GSF National Research Center for Environment and Health, Munich, Germany
Transcriptional regulation can occur at the level of initiation and
RNA elongation. We report that the rearranged, nontranscribed Ig
gene in the pre-B cell line 70Z/3 harbors a paused RNA polymerase II
(pol II) at a position between 45 and 89 bp downstream of the
transcription initiation site. LPS, an inducer of NF-B, activated
Ig gene transcription by increasing the processivity of pol II.
TGF-ß inhibited the LPS-induced transcription of the Ig gene, but
not initiation and pausing of pol II. A rearranged copy of the Ig
gene was introduced into 70Z/3 cells using an episomal vector system.
The episomal Ig was regulated by LPS and TGF-ß like the endogenous
gene and established a paused pol II, whereas a construct with a
deletion of the intron enhancer and the C region did not establish a
paused pol II. Two distinct functions can therefore be assigned to the
deleted DNA elements: loading of pol II to its pause site and induction
of processive transcription upon LPS stimulation. It had been proposed
that somatic hypermutation of Ig genes is connected to transcription.
The pause site of pol II described in this work resides upstream of the
previously defined 5' boundary of mutator activity at Ig genes. The
possible role of pausing of pol II for somatic hypermutation is
discussed.
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