A novel cell-cycle-indicator, mVenus-p27K-, identifies quiescent cells and visualizes G0-G1 transition

Toshihiko Oki; Koutarou Nishimura; Jiro Kitaura; Katsuhiro Togami; Akie Maehara; Kumi Izawa; Asako Sakaue-Sawano; Atsushi Niida; Satoru Miyano; Hiroyuki Aburatani; Hiroshi Kiyonari; Atsushi Miyawaki; Toshio Kitamura

doi:10.1038/srep04012

A novel cell-cycle-indicator, mVenus-p27K-, identifies quiescent cells and visualizes G0-G1 transition

Sci Rep. 2014 Feb 6:4:4012. doi: 10.1038/srep04012.

Affiliations

¹ 1] Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan [2] Division of Stem Cell Signaling, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
² Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
³ 1] Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN [2] Life Function and Dynamics, ERATO, JST, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
⁴ Laboratory of DNA Information Analysis, Human Genome Center, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
⁵ 1] Laboratory of DNA Information Analysis, Human Genome Center, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan [2] Laboratory of Sequence Analysis, Human Genome Center, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai Minato-ku, Tokyo 108-8639, Japan.
⁶ Genome Science Division, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
⁷ Laboratory for Animal Resources and Genetic Engineering, RIKEN Center for Developmental Biology, 2-2-3 Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan.

Abstract

The quiescent (G0) phase of the cell cycle is the reversible phase from which the cells exit from the cell cycle. Due to the difficulty of defining the G0 phase, quiescent cells have not been well characterized. In this study, a fusion protein consisting of mVenus and a defective mutant of CDK inhibitor, p27 (p27K(-)) was shown to be able to identify and isolate a population of quiescent cells and to effectively visualize the G0 to G1 transition. By comparing the expression profiles of the G0 and G1 cells defined by mVenus-p27K(-), we have identified molecular features of quiescent cells. Quiescence is also an important feature of many types of stem cells, and mVenus-p27K(-)-transgenic mice enabled the detection of the quiescent cells with muscle stem cell markers in muscle in vivo. The mVenus-p27K(-) probe could be useful in investigating stem cells as well as quiescent cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

3T3 Cells
Animals
Apoptosis Regulatory Proteins / biosynthesis
Apoptosis Regulatory Proteins / genetics
Bacterial Proteins / genetics*
Carrier Proteins / biosynthesis
Carrier Proteins / genetics
Cell Line
Cyclin-Dependent Kinase Inhibitor p16 / biosynthesis
Cyclin-Dependent Kinase Inhibitor p21 / biosynthesis
Cyclin-Dependent Kinase Inhibitor p27 / genetics*
Cyclin-Dependent Kinase Inhibitor p57 / biosynthesis
G1 Phase / genetics*
Gene Expression
Intracellular Signaling Peptides and Proteins
Luminescent Proteins / genetics*
Mice
Mice, Transgenic
MicroRNAs / biosynthesis
RNA-Binding Proteins / biosynthesis
RNA-Binding Proteins / genetics
Recombinant Fusion Proteins / biosynthesis
Recombinant Fusion Proteins / genetics*
Resting Phase, Cell Cycle / genetics*
Retinoblastoma Protein / biosynthesis
Selenium-Binding Proteins / biosynthesis
Selenium-Binding Proteins / genetics
Stem Cells / cytology
Tumor Suppressor Protein p53 / biosynthesis

Substances

Apoptosis Regulatory Proteins
Bacterial Proteins
Carrier Proteins
Cdkn1a protein, mouse
Cdkn1c protein, mouse
Cdkn2a protein, mouse
Cyclin-Dependent Kinase Inhibitor p16
Cyclin-Dependent Kinase Inhibitor p21
Cyclin-Dependent Kinase Inhibitor p57
Intracellular Signaling Peptides and Proteins
Luminescent Proteins
MicroRNAs
Pdcd4 protein, mouse
RNA-Binding Proteins
Recombinant Fusion Proteins
Retinoblastoma Protein
Selenbp1 protein, mouse
Selenium-Binding Proteins
Tob1 protein, mouse
Tumor Suppressor Protein p53
mirnlet7 microRNA, mouse
yellow fluorescent protein, Bacteria
Cyclin-Dependent Kinase Inhibitor p27