The formation of eukaryotic mRNAs involves the cleavage and polyadenylation of pre- mRNAs. To investigate the sequence requirement of putative polyadenylation signals (PASs), poly(A) sites and downstream elements (DUEs) in 3′-end-pro- cessing in rice, we compared expressed sequences tags (ESTs) with poly(A) extremity to full-length cDNA sequences and constructed a database of 12969 pre- mRNA sequences in ?40―+40 nt surrounding the poly(A) sites, which were from 9953 genes. The al- ternative poly(A) sites were revealed in approxi- mately 25% of mRNAs. Nearly 80% of pre-mRNAs showed stringent requirement of the YA (CA or UA) at poly (A) sites for polyadenylation. About 7.9% had the AAUAAA signals on ?40―?1 nt upstream of the poly(A) sites. Over 60% of mRNAs probably used the one- or two-base variants of AAUAAA hexamers as their PASs in 3′ fragments. The single-base variants of AAUGAA revealed the high frequency in 11.5% of 3′ fragments. The DUEs were detected in 90% of pre- mRNAs, especially more than half of the pre-mRNAs with multi-base variants of AAUAAA had the DUEs surrounding the poly(A) site. The location of DUE is also important for defining the cleavage site. Al- though most of the rice pre-mRNAs did not contain AAUAAA signal, the existence of downstream ele- ments ensured the efficiency of cleavage-polyade- nylation The formation of eukaryotic mRNAs involves the cleavage and polyadenylation of pre-mRNAs. To investigate the sequence requirement of putative polyadenylation signals (PASs), poly (A) sites and downstream elements (DUEs) in 3’-end-pro-cessing in rice , we were expressing sequences tags (ESTs) with poly (A) extremity to full-length cDNA sequences and constructed a database of 12969 pre- mRNA sequences in? 40- + 40 nt surrounding the poly (A) sites, which were from 9953 genes. The al- ternative poly (A) sites were revealed in approxi- mately 25% of mRNAs. Nearly 80% of pre-mRNAs displayed stringent requirement of the YA (CA or UA) at poly (A) sites for polyadenylation. 7.9% had the AAUAAA signals on? 40-? 1 nt upstream of the poly (A) sites. Over 60% of mRNAs probably used the one- or two-base variants of AAUAAA hexamers as their PASs in 3 ’fragments. The single -base variants of AAUGAA revealed the high frequency in 11.5% of 3 ’fragments. The DUEs were detected in 90% of pre- mRN As, especially more than half of the pre-mRNAs with multi-base variants of AAUAAA had the DUEs surrounding the poly (A) site. The location of DUE is also important for defining the cleavage site. Al- though most of the rice pre -mRNAs did not contain AAUAAA signal, the existence of downstream ele ments won the efficiency of cleavage-polyade- nylation