Molecular cloning and characterization of profilin from tobacco (Nicotiana tabacum): increased profilin expression during pollen maturation.
Profilin has recently been identified as an actin-binding protein in higher plants. A cDNA coding for tobacco profilin, which shared an average sequence identity of 75% with other plant profilins, was isolated from a tobacco pollen cDNA library by antibody screening. Tobacco profilin was expressed in Escherichia coli and purified by affinity to poly-(L-proline) Sepharose. A rabbit antiserum was raised against recombinant tobacco profilin and used to estimate the amount of profilin expressed in different tobacco tissues. Profilin can be detected in different somatic tissues, but the expression is 50-100 fold higher in mature pollen. Immunofluorescence and confocal laser scanning microscopy showed a homogeneous distribution of profilin in the cytoplasm of in vitro cultured pollen grains and pollen tubes of tobacco whereas some growing pollen tubes were stained more intensively a their tip. A possible role of pollen profilin as a developmentally upregulated microfilament precursor in mature pollen is discussed.
Mittermann
I
I
Division of Immunopathology, AKH, University of Vienna, Austria.
Swoboda
I
I
Pierson
E
E
Eller
N
N
Kraft
D
D
Valenta
R
R
Heberle-Bors
E
E
D000199:Actins / Q000032:analysis; D000595:Amino Acid Sequence; D015152:Blotting, Northern; D002451:Cell Compartmentation; D003285:Contractile Proteins; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015723:Gene Library; D015151:Immunoblotting; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D010947:Plants, Toxic; D011058:Pollen / Q000254:growth & development* / Q000648:ultrastructure; D051304:Profilins; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014018:Tissue Distribution; D014026:Nicotiana / Q000235:genetics* / Q000254:growth & development
D000199:Actins; D003285:Contractile Proteins; D018076:DNA, Complementary; D008840:Microfilament Proteins; D010940:Plant Proteins; D051304:Profilins; D011994:Recombinant Proteins