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Molecular and Cellular Biology, March 2001, p. 1509-1514, Vol. 21, No. 5
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.5.1509-1514.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

An Upstream AG Determines Whether a Downstream AG Is Selected during Catalytic Step II of Splicing

Katrin Chua and Robin Reed*

Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115

Received 31 July 2000/Returned for modification 14 September 2000/Accepted 5 December 2000

Specific mechanisms must exist to ensure fidelity in selecting the AG dinucleotide that functions as the 3' splice site during the second transesterification step of splicing. Here we show that the optimal location for this AG is within a narrow distance (19 to 23 nucleotides [nt]) downstream from the branch point sequence (BPS). Contrary to previous expectations, AGs located less than 23 nt from the BPS are always recognized, even when a second AG located more optimally downstream is used in the transesterification reaction. Indeed, the AG closest to the BPS actually dictates the precise location of the AG that engages in the reaction. This mechanism, in which the AG is identified by a general localization step followed by a precise localization step, may be used to achieve fidelity while allowing flexibility in the location of 3' splice sites.

* Corresponding author. Mailing address: Department of Cell Biology, Harvard Medical School, Boston, MA 02115. Phone: (617) 432-2844. Fax: (617) 432-3091. E-mail: rreed{at}hms.harvard.edu.

Molecular and Cellular Biology, March 2001, p. 1509-1514, Vol. 21, No. 5
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.5.1509-1514.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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