Characterization of SP1, a stress-responsive, boiling-soluble, homo-oligomeric protein from aspen.
sp1 cDNA was isolated from aspen (Populus tremula) plants by immunoscreening an expression library using polyclonal antibodies against BspA protein. BspA, which is a boiling-stable protein, accumulates in aspen plants in response to water stress and abscisic acid application (Pelah et al., 1995). The sp1 cDNA was found to encode a 12.4-kD generally hydrophilic protein with a hydrophobic C terminus, which is different from the BspA protein and was termed SP1 (stable protein 1). Northern-blot analysis revealed that sp1 encodes a small mRNA (about 0.6 kb) that is expressed in aspen plants under non-stress conditions and is accumulated after salt, cold, heat, and desiccation stress, and during the recovery from stress. The SP1 detected in plants remained soluble upon boiling, migrated both as a 12.4-kD band and a much higher mass of 116 kD on a 17% (w/v) Tricine-sodium dodecyl sulfate-polyacrylamide gel. Comparative protease digestion patterns, amino acid analyses, and the N-terminal sequences of the 12.4- and 116-kD proteins revealed that SP1 is homo-oligomeric. Furthermore, gel filtration chromatography analysis indicated that SP1 exists in aspen plants as a complex, composed of 12 subunits of 12.4 kD. A large number of sequences deduced from expressed sequence tags and genomic sequences of other organisms with unknown function show high homology to SP1. Thus, SP1 may represent a new protein family. Here, we present the first report on this putative protein family: the cloning, isolation, and characterization of SP1, a stress-responsive, boiling-soluble, oligomeric protein.
Wang
Wang-Xia
WX
The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel.
Pelah
Dan
D
Alergand
Tal
T
Shoseyov
Oded
O
Altman
Arie
A
D000040:Abscisic Acid / Q000494:pharmacology; D000222:Adaptation, Physiological / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003513:Cycloheximide / Q000494:pharmacology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D008353:Mannitol / Q000494:pharmacology; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D032107:Populus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012965:Sodium Chloride / Q000494:pharmacology; D014867:Water / Q000494:pharmacology / Q000502:physiology
D018076:DNA, Complementary; D010940:Plant Proteins; D014867:Water; D008353:Mannitol; D012965:Sodium Chloride; D000040:Abscisic Acid; D003513:Cycloheximide