DNA-directed termination of RNA polymerase II transcription
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DNA-directed termination of RNA polymerase II transcription. / Han, Zhong; Moore, George A.; Mitter, Richard; Lopez Martinez, David; Wan, Li; Dirac Svejstrup, A. Barbara; Rueda, David S.; Svejstrup, Jesper Q.
In: Molecular Cell, Vol. 83, No. 18, 2023, p. 3253-3267.e7.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - DNA-directed termination of RNA polymerase II transcription
AU - Han, Zhong
AU - Moore, George A.
AU - Mitter, Richard
AU - Lopez Martinez, David
AU - Wan, Li
AU - Dirac Svejstrup, A. Barbara
AU - Rueda, David S.
AU - Svejstrup, Jesper Q.
N1 - Publisher Copyright: © 2023 The Authors
PY - 2023
Y1 - 2023
N2 - RNA polymerase II (RNAPII) transcription involves initiation from a promoter, transcriptional elongation through the gene, and termination in the terminator region. In bacteria, terminators often contain specific DNA elements provoking polymerase dissociation, but RNAPII transcription termination is thought to be driven entirely by protein co-factors. We used biochemical reconstitution, single-molecule studies, and genome-wide analysis in yeast to study RNAPII termination. Transcription into natural terminators by pure RNAPII results in spontaneous termination at specific sequences containing T-tracts. Single-molecule analysis indicates that termination involves pausing without backtracking. The “torpedo” Rat1-Rai1 exonuclease (XRN2 in humans) greatly stimulates spontaneous termination but is ineffectual on other paused RNAPIIs. By contrast, elongation factor Spt4-Spt5 (DSIF) suppresses termination. Genome-wide analysis further indicates that termination occurs by transcript cleavage at the poly(A) site exposing a new 5′ RNA-end that allows Rat1-Rai1 loading, which then catches up with destabilized RNAPII at specific termination sites to end transcription.
AB - RNA polymerase II (RNAPII) transcription involves initiation from a promoter, transcriptional elongation through the gene, and termination in the terminator region. In bacteria, terminators often contain specific DNA elements provoking polymerase dissociation, but RNAPII transcription termination is thought to be driven entirely by protein co-factors. We used biochemical reconstitution, single-molecule studies, and genome-wide analysis in yeast to study RNAPII termination. Transcription into natural terminators by pure RNAPII results in spontaneous termination at specific sequences containing T-tracts. Single-molecule analysis indicates that termination involves pausing without backtracking. The “torpedo” Rat1-Rai1 exonuclease (XRN2 in humans) greatly stimulates spontaneous termination but is ineffectual on other paused RNAPIIs. By contrast, elongation factor Spt4-Spt5 (DSIF) suppresses termination. Genome-wide analysis further indicates that termination occurs by transcript cleavage at the poly(A) site exposing a new 5′ RNA-end that allows Rat1-Rai1 loading, which then catches up with destabilized RNAPII at specific termination sites to end transcription.
KW - CPSF73
KW - DSIF
KW - intrinsic termination site
KW - Rat1
KW - RNA polymerase II
KW - Spt5
KW - termination
KW - TFIIS
KW - torpedo
KW - XRN2
U2 - 10.1016/j.molcel.2023.08.007
DO - 10.1016/j.molcel.2023.08.007
M3 - Journal article
C2 - 37683646
AN - SCOPUS:85171165345
VL - 83
SP - 3253-3267.e7
JO - Molecular Cell
JF - Molecular Cell
SN - 1097-2765
IS - 18
ER -
ID: 373468088