(A) Sequence identities at the 5′ ends of cDNA clones
from Hydra genes. Identical 5′ sequences are in
lowercase; divergent downstream sequences are in uppercase. The
translation start ATG codon is separated from the 5′ untranslated
region sequence by a slash. The upper group of sequences contains the
spliced leader A (SL-A) sequence; the lower group contains the spliced
leader B (SL-B) sequence. GenBank accession nos. for the sequences are
as follows: Hint, M64611; Syk, AF060949; HTK32,
AF123442; HTK54, U24116; HFZ, AF209200; Csk, AF067775;
Cnash, U36275; Alx, AF295531; Pax-A, U96193; enolase,
U85827; PLC-βHI, AB017511; hyGK, AF031931; ECE, AF162671; hym-323,
AB40074; HTK16, U00936; HZO-1, AF230482; PKC1B, Y12857; ras1, X78597;
nucleoporin, U85827; annexin XII, M83736. All genes are from H.
vulgaris except hyGK (Hydra oligactis), enolase
(H. oligactis), Pax-A (Hydra littoralis),
and PLC-βI (Hydra magnipapillata). The three different
Hint sequences (labeled Hint 1–3) arise because of
alternative splicing. Hint produces a long transcript
with SL-A at the 5′ end and a shorter transcript that can contain
either SL-A or -B (45). The single nucleotide difference (T>G) in the
SL-A sequence of HTK54 may be because of an error during cDNA synthesis
or the presence of multiple alleles of SL-A. A complete copy of the
SL-B sequence is located internally in a H. vulgaris
cDNA for cAMP-response element-binding protein (CREB) (46), where it
results in the truncation of a highly conserved portion of the CREB
coding sequence. We have attempted, without success, to confirm this
arrangement by amplification of the corresponding region from
first-strand cDNA made from H. vulgaris polyA+ RNA. We
therefore believe that this clone is a hybrid produced during cDNA
library construction by ligation of the 3′ end of a partial CREB cDNA
to the 5′ end of a cDNA derived from an SL-B-containing mRNA.
(B) Alignment of sequences from the 5′ ends of HTK32
cDNA clones containing SL-A or -B sequences. The clone containing SL-B
includes four nucleotides that are not present in the SL-A-containing
message. The splice acceptor dinucleotide is underlined. The
SL-A-containing sequence is from a clone isolated from a cDNA library.
The SL-B-containing sequence was obtained by 5′ RACE (see
Materials and Methods). (C) Genomic
sequence from the Hydra Syk gene (24). The splice
acceptor dinucleotide is indicated by double underlining. The genomic
sequence is aligned with the sequences from Syk cDNAs containing either
SL-A or -B (see A). The pyrimidine-rich sequence
upstream of the splice acceptor dinucleotide is shaded.