PMID- 2837493
OWN - NLM
STAT- MEDLINE
DCOM- 19880727
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 263
IP  - 19
DP  - 1988 Jul 5
TI  - A role for divalent cations in specifying the start site for transcription from 
      chromatin templates in vitro.
PG  - 9550-6
AB  - An assay that employs nucleoside 5'-O-(2-thiotriphosphates) was used to detect 
      initiation of mouse mammary tumor virus (MMTV) RNA chains in preparations of 
      isolated nuclei from cultured rat hepatoma cells containing stably integrated 
      proviruses. RNA chains initiated with adenosine 5'-O-(2-thiotriphosphate), 
      guanosine 5'-O-(2-thiotriphosphate), or uridine 5'-O-(2-thiotriphosphate) were 
      separated from the remaining RNA by mercury-Sepharose column chromatography and 
      analyzed for correctly initiated RNA chains with a T1 nuclease protection assay. 
      Combined use of the thionucleotide transcription reaction with the T1 nuclease 
      assay allowed precise localization of the transcription start sites. The majority 
      of MMTV RNA chains were initiated with guanosine 5'-O-(2-thiotriphosphate) at a 
      template site 133 nucleotides upstream from a PvuII site that coincides with the 
      right end of the long terminal repeat. However, some RNA chains were also 
      initiated with adenosine 5'-O-(2-thiotriphosphate) and uridine 
      5'-O-(2-thiotriphosphate) at template sites within three nucleotides of the 
      primary guanosine start site. When Mn2+ was substituted for Mg2+ in the 
      transcription reaction, MMTV RNA chains were initiated with approximately the 
      same efficiency, but the start site was shifted to a position approximately 40 
      nucleotides downstream from the physiological start site; in the presence of 
      Mn2+, MMTV RNA chains were initiated only with guanosine 
      5'-O-(2-thiotriphosphate). When the nuclei were exposed to both Mn2+ and Mg2+, 
      transcription initiated at the manganese-dependent site. Mn2+ also caused the 
      transcription start site for 45 S pre-rRNA to shift about 10 nucleotides upstream 
      from the physiologically correct start site.
FAU - Zhang-Keck, Z Y
AU  - Zhang-Keck ZY
AD  - Department of Pathology, University of Southern California School of Medicine, 
      Los Angeles 90033.
FAU - Eckstein, F
AU  - Eckstein F
FAU - Washington, L D
AU  - Washington LD
FAU - Stallcup, M R
AU  - Stallcup MR
LA  - eng
GR  - CA01147/CA/NCI NIH HHS/United States
GR  - GM36317/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Cations, Divalent)
RN  - 0 (Chromatin)
RN  - 0 (RNA, Viral)
RN  - 0 (Thionucleotides)
RN  - 37589-80-3 (Guanosine 5'-O-(3-Thiotriphosphate))
RN  - 42Z2K6ZL8P (Manganese)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - I38ZP9992A (Magnesium)
SB  - IM
MH  - Animals
MH  - Cations, Divalent
MH  - Cell Line, Transformed
MH  - Cell Nucleus/metabolism
MH  - Chromatin/drug effects/*metabolism
MH  - Guanosine 5'-O-(3-Thiotriphosphate)
MH  - Guanosine Triphosphate/analogs & derivatives/pharmacology
MH  - Liver Neoplasms, Experimental
MH  - Magnesium/*pharmacology
MH  - Mammary Tumor Virus, Mouse/genetics
MH  - Manganese/*pharmacology
MH  - Nucleic Acid Hybridization
MH  - RNA, Viral/genetics
MH  - Templates, Genetic
MH  - Thionucleotides/pharmacology
MH  - Transcription, Genetic/*drug effects
EDAT- 1988/07/05 00:00
MHDA- 1988/07/05 00:01
CRDT- 1988/07/05 00:00
PHST- 1988/07/05 00:00 [pubmed]
PHST- 1988/07/05 00:01 [medline]
PHST- 1988/07/05 00:00 [entrez]
AID - S0021-9258(19)76577-6 [pii]
PST - ppublish
SO  - J Biol Chem. 1988 Jul 5;263(19):9550-6.