The molecular and biochemical characterization of cell cycle regulators in Pisum sativum
Feiler, Heidi Sue
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https://hdl.handle.net/2142/22279
Description
Title
The molecular and biochemical characterization of cell cycle regulators in Pisum sativum
Author(s)
Feiler, Heidi Sue
Issue Date
1992
Doctoral Committee Chair(s)
Jacobs, Thomas W.
Department of Study
Plant Biology
Discipline
Plant Biology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Biology, Botany
Chemistry, Biochemistry
Language
eng
Abstract
This dissertation focuses on the characterization of cdc2-type cell cycle regulators in Pisum sativum (garden pea). Immunoblot patterns indicate that $\sim$34 kDa cdc2-type proteins present in protein complexes may be phosphorylated in pea. As in other systems, these proteins copurify with a histone H1 kinase activity by p13$\sp{suc1}$-Sepharose affinity chromatography. p13$\sp{suc1}$-associated histone H1 kinase activity is elevated and a set of endogenous proteins is phosphorylated specifically at the S and M phases. Cell cycle changes in the p34 immunoblot pattern are not detected. These results suggest that, as in other systems, a cascade of protein kinases may regulate the plant cell cycle.
Segments of a putative cdc2 gene (cdc2Ps), a cdc2-like gene (cdc2Ps-like) and a cyclin gene (cyclin Ps 6-8) were isolated from pea cDNA by polymerase chain reaction (PCR). cdc2Ps encodes all 16 amino acids of the PSTAIRE domain characteristic of every cyclin-dependent kinase (CDK) gene product sequence examined to date. The derived amino acid sequence of the cdc2Ps PCR product is 87-95% identical to the other plant cdc2 homologues. These results suggest that cdc2Ps is a segment of a pea CDK gene, and most likely a cdc2 homologue. cdc2Ps-like (designated pcl1) is not a member of the CDK family, as it encodes only 11 out of the 16 amino acids of the PSTAIRE domain and appears most closely related to a human cdc2-like kinase (HUMCHED). The derived amino acid sequence of the cyclinPs 6-8 (designated psc1) PCR product is 85% identical to a soybean cyclin (SOYMCYC).
The research presented here indicates that CDKs and cyclins are synthesized in higher plants and suggests that CDK regulation functions as the basic cell cycle control network in higher plants.
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