Four distinct and unusual linker proteins in a mitotically dividing nucleus are derived from a 71-kilodalton polyprotein, lack p34cdc2 sites, and contain protein kinase A sites.
Tetrahymena thermophila micronuclei contain four linker-associated proteins, alpha, beta, gamma, and delta. Synthetic oligonucleotides based on N-terminal protein sequences of beta and gamma were used to clone the micronuclear linker histone (MLH) gene. The MLH gene is single copy and is transcribed into a 2.4-kb message encoding all four linker-associated proteins. The message is translated into a polypeptide (Mic LH) that is processed at the sequence decreases RTK to give proteins whose amino acid sequences differ markedly from each other, from the sequence of macronuclear H1, and from sequences of typical H1s of other organisms. This represents the first example of multiple chromatin proteins derived from a single polyprotein. The delta protein consists largely of two high-mobility-group (HMG) boxes. An evolutionary analysis of HMG boxes indicates that the delta HMG boxes are similar to the HMG boxes of tsHMG, a protein that appears in elongating mouse spermatids when they condense and cease transcription, suggesting that delta could play a similar role in the micronucleus. The micronucleus divides mitotically, while the macronucleus divides amitotically. Surprisingly, macronuclear H1 but not Mic LH contains sequences resembling p34cdc2 kinase phosphorylation sites, while each of the Mic LH-derived proteins contains a typical protein kinase A phosphorylation site in its carboxy terminus.
Wu
M
M
Department of Biology, University of Rochester, New York 14627.
Allis
C D
CD
Sweet
M T
MT
Cook
R G
RG
Thatcher
T H
TH
Gorovsky
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000378:metabolism; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D016054:DNA, Protozoan / Q000235:genetics; D017125:Genes, Protozoan; D006609:High Mobility Group Proteins / Q000235:genetics; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D048631:Micronucleus, Germline / Q000378:metabolism; D008938:Mitosis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D010802:Phylogeny; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D016808:Tetrahymena thermophila / Q000166:cytology / Q000235:genetics / Q000378:metabolism*
D016054:DNA, Protozoan; D006609:High Mobility Group Proteins; D006657:Histones; D015800:Protozoan Proteins; D017868:Cyclic AMP-Dependent Protein Kinases; D016203:CDC2 Protein Kinase