title: Purification and characterization of poly(A) polymerase from Saccharomyces cerevisiae.
abstract: Poly(A) polymerase was purified 22,000-fold to homogeneity from a whole cell extract of Saccharomyces cerevisiae with a yield of 22%. The enzyme is a monomeric polypeptide with a denatured molecular weight of 63,000. Incorporation of labeled ATP into acid-precipitable material by the purified enzyme proceeds faster with manganese than with magnesium ions. Various RNA homopolymers as well as Escherichia coli tRNA or rRNA can serve as primers. An RNA that terminates at the natural poly(A) site of the CYC1 gene is not more efficiently elongated than several nonspecific substrates, indicating the requirement for additional factors to provide specificity. Elongation of the primer is distributive. Covering of a poly(A) primer with poly(A)-binding protein reduces the enzyme's activity more than 10-fold.
authors.affiliation: Department of Cell Biology, University of Basel, Switzerland.
authors.forename: J
authors.initials: J
authors.lastname: Lingner
authors.forename: I
authors.initials: I
authors.lastname: Radtke
authors.forename: E
authors.initials: E
authors.lastname: Wahle
authors.forename: W
authors.initials: W
authors.lastname: Keller
mesh_terms: D002352:Carrier Proteins / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D011061:Poly A / Q000378:metabolism; D039102:Poly(A)-Binding Proteins; D011062:Polynucleotide Adenylyltransferase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
chemical_list: D002352:Carrier Proteins; D005656:Fungal Proteins; D039102:Poly(A)-Binding Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D011061:Poly A; D011062:Polynucleotide Adenylyltransferase