HORMONAL-REGULATION, PROCESSING, AND SECRETION OF CYSTEINE PROTEINASES IN BARLEY ALEURONE LAYERS

被引:154
作者
KOEHLER, SM [1 ]
HO, THD [1 ]
机构
[1] WASHINGTON UNIV,DEPT BIOL,ST LOUIS,MO 63130
关键词
D O I
10.1105/tpc.2.8.769
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Barley aleurone layers synthesize and secrete several proteases in response to gibberellic acid (GA3). Two major cysteine proteinases designated EP-A (37,000 Mr) and EP-B (30,000 Mr) have been described [Koehler and Ho (1988). Plant Physiol. 87, 95-103]. We now report the cDNA cloning of EP-B and describe the post-translational processing and hormonal regulation of both cysteine proteinases. Three cDNAs for cysteine proteinases were cloned from GA3-induced barley aleurone layers. Genomic DNA gel blot analysis indicated that these are members of a small gene family with no more than four to five different genes. The proteins encoded by two of these clones, pHVEP1 and 4, are 98% similar to each other and are isozymes of EP-B. The proteins contain large preprosequences followed by the amino acid sequence described as the mature N terminus of purified EP-B, and are antigenic to EP-B antiserum. The results of pulse-chase experiments indicated that the post-translational processing of large prosequences proceeds in a multistep fashion to produce the mature enzymes. Processing intermediates for EP-B are observed both in the aleurone layers and surrounding incubation medium, but only mature EP-A is secreted. The regulation of synthesis of EP-A, EP-B, and other aleurone cysteine proteinases was compared at the protein and mRNA levels. We conclude that barley aleurone cysteine proteinases are differentially regulated with respect to their temporal and hormonally induced expression.
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页码:769 / 783
页数:15
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共 50 条
[1]  
AKASOFU H, 1989, NUCLEIC ACIDS RES, V17, P3733
[2]   STRUCTURE OF ACTINIDIN, AFTER REFINEMENT AT 1.7-A RESOLUTION [J].
BAKER, EN .
JOURNAL OF MOLECULAR BIOLOGY, 1980, 141 (04) :441-484
[3]   HEAT-SHOCK CAUSES DESTABILIZATION OF SPECIFIC MESSENGER-RNAS AND DESTRUCTION OF ENDOPLASMIC-RETICULUM IN BARLEY ALEURONE CELLS [J].
BELANGER, FC ;
BRODL, MR ;
HO, TD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (05) :1354-1358
[4]   ANTIBODY TO TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE [J].
BOLLUM, FJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1975, 72 (10) :4119-4122
[5]   AMINO-ACID SEQUENCE OF TRYPTIC PEPTIDES FROM ACTINIDIN, A PROTEOLYTIC-ENZYME FROM FRUIT OF ACTINIDIA-CHINESIS [J].
CARNE, A ;
MOORE, CH .
BIOCHEMICAL JOURNAL, 1978, 173 (01) :73-83
[6]   NUCLEOTIDE AND PREDICTED AMINO-ACID-SEQUENCES OF CLONED HUMAN AND MOUSE PREPROCATHEPSIN-B CDNAS [J].
CHAN, SJ ;
SANSEGUNDO, B ;
MCCORMICK, MB ;
STEINER, DF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (20) :7721-7725
[7]   THE EFFECTS OF GIBBERELLIC-ACID AND ABSCISIC-ACID ON ALPHA-AMYLASE MESSENGER-RNA LEVELS IN BARLEY ALEURONE LAYERS STUDIES USING AN ALPHA-AMYLASE CDNA CLONE [J].
CHANDLER, PM ;
ZWAR, JA ;
JACOBSEN, JV ;
HIGGINS, TJV ;
INGLIS, AS .
PLANT MOLECULAR BIOLOGY, 1984, 3 (06) :407-418
[8]   GENOMIC SEQUENCING [J].
CHURCH, GM ;
GILBERT, W .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (07) :1991-1995
[9]  
COHEN LW, 1986, GENE, V48, P219, DOI 10.1016/0378-1119(86)90080-6
[10]   A COMPREHENSIVE SET OF SEQUENCE-ANALYSIS PROGRAMS FOR THE VAX [J].
DEVEREUX, J ;
HAEBERLI, P ;
SMITHIES, O .
NUCLEIC ACIDS RESEARCH, 1984, 12 (01) :387-395