Reference "Coordinate increase in major transcripts from the high pI alpha-amylase multigene family in barley aleurone cells stimulated with gibberellic acid"

Reference ID

8826

Title

Coordinate increase in major transcripts from the high pI alpha-amylase multigene family in barley aleurone cells stimulated with gibberellic acid

Source

The Journal of biological chemistry, 1984, vol. 259, pp. 12234-12240

Authors (2)

Rogers-J-C

Milliman-C

Abstract

The purpose of this study was to identify specifically genes and transcripts
for the high pI isozyme of barley alpha-amylase. From hybridization of coding
sequence probes to blots of genomic DNA digested with restriction enzymes that
do not cut within our cloned high pI alpha-amylase cDNA, it is estimated that
about 7 alpha-amylase genes or pseudogenes exist. No difference could be
detected between barley aleurone cell and sprout DNAs. Experiments using
probes from the 5' and 3' untranslated sequences of the high pI alpha-amylase
cDNA clone identified three HindIII fragments that probably carry high pI
sequences. Primer extension experiments used as a primer the terminal 5'
coding sequence from our cDNA clone; this primer would not cross-hybridize to
low pI alpha-amylase transcripts. Two major transcripts were identified. These
shared a conserved 23-base sequence immediately 5' to the ATG start codon,
although a C----G transversion and a 3-base deletion were present within this
sequence. An unusual 8-base pair GC palindrome was present in the conserved
region immediately preceding the ATG start codon. Distal to the conserved
sequence there was no apparent homology. One transcript carrying a 97-base
untranslated region was identical to our high pI cDNA clone E. The gene for
the other was recovered from a lambda phage genomic library. The 5' coding
sequence was very similar, but not identical to clone E, demonstrating that
these transcripts arise from separate genes. The two transcripts increased
coordinately in aleurone cells stimulated with gibberellic acid. These data
indicate that there is a high pI alpha-amylase multigene family with at least
two active members, both of which are regulated in some manner by the plant
hormone gibberellic acid.