Light-modulated abundance of an mRNA encoding a calmodulin-regulated, chromatin-associated NTPase in pea.
A CDNA encoding a 47 kDa nucleoside triphosphatase (NTPase) that is associated with the chromatin of pea nuclei has been cloned and sequenced. The translated sequence of the cDNA includes several domains predicted by known biochemical properties of the enzyme, including five motifs characteristic of the ATP-binding domain of many proteins, several potential casein kinase II phosphorylation sites, a helix-turn-helix region characteristic of DNA-binding proteins, and a potential calmodulin-binding domain. The deduced primary structure also includes an N-terminal sequence that is a predicted signal peptide and an internal sequence that could serve as a bipartite-type nuclear localization signal. Both in situ immunocytochemistry of pea plumules and immunoblots of purified cell fractions indicate that most of the immunodetectable NTPase is within the nucleus, a compartment proteins typically reach through nuclear pores rather than through the endoplasmic reticulum pathway. The translated sequence has some similarity to that of human lamin C, but not high enough to account for the earlier observation that IgG against human lamin C binds to the NTPase in immunoblots. Northern blot analysis shows that the NTPase MRNA is strongly expressed in etiolated plumules, but only poorly or not at all in the leaf and stem tissues of light-grown plants. Accumulation of NTPase mRNA in etiolated seedlings is stimulated by brief treatments with both red and far-red light, as is characteristic of very low-fluence phytochrome responses. Southern blotting with pea genomic DNA indicates the NTPase is likely to be encoded by a single gene.
Hsieh
H L
HL
Department of Botany, The University of Texas at Austin, 78713, USA.
Tong
C G
CG
Thomas
C
C
Roux
S J
SJ
D017766:Acid Anhydride Hydrolases / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002147:Calmodulin / Q000378:metabolism*; D002843:Chromatin / Q000201:enzymology; D018506:Gene Expression Regulation, Plant / Q000528:radiation effects*; D015723:Gene Library; D015151:Immunoblotting; D008027:Light; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D043583:Nucleoside-Triphosphatase; D018532:Pisum sativum / Q000235:genetics* / Q000528:radiation effects; D011487:Protein Conformation; D012333:RNA, Messenger / Q000096:biosynthesis; D018749:RNA, Plant / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
D002147:Calmodulin; D002843:Chromatin; D012333:RNA, Messenger; D018749:RNA, Plant; D017766:Acid Anhydride Hydrolases; D043583:Nucleoside-Triphosphatase