Cloning of a cDNA encoding a developmentally regulated 22 kDa polypeptide from tobacco leaf plasma membrane.
A polypeptide doublet (P18-P19, ca 22 kDa, pI 4.5) has been shown to accumulate in tobacco leaf plasma membrane in a development-dependent way, under constant environmental conditions. P18 and P19 were purified by 2D-PAGE and microsequenced. Microsequences revealed only small differences between the two polypeptides. A PCR-based cloning strategy identified a cDNA displaying a 591 bp ORF. The encoded polypeptide contained P19 specific microsequences. It was expressed in E. coli and a specific rabbit antiserum was raised. Western-blots confirmed its identification as P19. The accumulation pattern of hybridizable mRNA around the floral induction period was similar to that of P18 and P19. Searching of databases revealed no significant hits except unidentified plant ESTs. P18 and P19 are proposed as the first example of plant-specific and developmentally regulated plasma membrane proteins.
Gantet
P
P
INRA/ENSA-M/CNRS URA 573, Laboratoire de Biochimie et Physiologie Végétales, Montpellier, France.
Masson
F
F
Domergue
O
O
Marquis-Mention
M
M
Bauw
G
G
Inze
D
D
Rossignol
M
M
de la Serve
B T
BT
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017403:In Situ Hybridization; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010452:Peptide Biosynthesis; D010455:Peptides / Q000235:genetics* / Q000276:immunology; D018515:Plant Leaves / Q000737:chemistry / Q000235:genetics*; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000235:genetics* / Q000254:growth & development / Q000502:physiology
D000906:Antibodies; D018076:DNA, Complementary; D008565:Membrane Proteins; D010455:Peptides; D010940:Plant Proteins; D012333:RNA, Messenger