Stability of the replicative Mcm3 protein in proliferating and differentiating human cells.
Mcm proteins are abundant nuclear proteins involved in the regulation of genome replication. Previous experiments had shown that levels of Mcm-specific mRNAs increase at the G1/S phase transition of the cell cycle, but that the amounts of Mcm proteins do not change much during the cell cycle. To learn more about the stability of an Mcm protein we performed experiments which showed that: (i) more than 60% of [35S]methionine pulse-labeled Mcm3 protein appears to be degraded during a 24-h chase in HeLa cells; (ii) the amount of Mcm3 protein significantly decreases during the differentiation of HL60 cells in vitro (whereas another replication-initiation protein, hOrc2, remains fairly constant); and (iii) according to immunohistochemical staining, Mcm3 protein is present in nuclei of cells in the proliferating zone of human epidermal tissue, but in decreasing amounts in nuclei of differentiating cells of the upper cell layers. Our interpretation is that Mcm3 protein is no longer synthesized after initiation of differentiation and slowly disappears at a half-life of approximately 24 h.
Musahl
C
C
Department of Biology, Universität Konstanz, Germany.
Holthoff
H P
HP
Lesch
R
R
Knippers
R
R
D018797:Cell Cycle Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D004268:DNA-Binding Proteins; D018922:HL-60 Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D006367:HeLa Cells / Q000166:cytology*; D006801:Humans; D008715:Methionine / Q000378:metabolism; D064126:Minichromosome Maintenance Complex Component 3; D009687:Nuclear Proteins; D012867:Skin / Q000737:chemistry / Q000166:cytology; D013462:Sulfur Radioisotopes
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C066470:MCM3 protein, human; D009687:Nuclear Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine; D064126:Minichromosome Maintenance Complex Component 3