Using single-particle tracking to study nuclear trafficking of viral genes

被引:108
作者
Babcock, HP
Chen, C
Zhuang, XW [1 ]
机构
[1] Harvard Univ, Dept Phys, Cambridge, MA 02138 USA
[2] Harvard Univ, Dept Chem & Biol Chem, Cambridge, MA 02138 USA
基金
美国国家科学基金会;
关键词
D O I
10.1529/biophysj.104.042234
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The question of how genetic materials are trafficked in and out of the cell nucleus is a problem of great importance not only for understanding viral infections but also for advancing gene-delivery technology. Here we demonstrate a physical technique that allows gene trafficking to be studied at the single-gene level by combining sensitive fluorescence microscopy with microinjection. As a model system, we investigate the nuclear import of influenza genes, in the form of ribonucleoproteins (vRNPs), by imaging single vRNPs in living cells in real time. Our single-particle trajectories show that vRNPs are transported to the nuclear envelope by diffusion. We have observed heterogeneous interactions between the vRNPs and nuclear pore complexes with dissociation rate constants spanning two orders of magnitude. Our single-particle tracking experiments also provided new insights into the regulation mechanisms for the nuclear import of vRNPs: the influenza M1 protein, a regulatory protein for the import process, downregulates the nuclear import of vRNPs by inhibiting the interactions between vRNPs and nuclear pore complexes but has no significant effect on the transport properties of vRNPs. We expect this single-particle tracking approach to find broad application in investigations of genetic trafficking.
引用
收藏
页码:2749 / 2758
页数:10
相关论文
共 46 条
  • [1] Role of the influenza virus M1 protein in nuclear export of viral ribonucleoproteins
    Bui, M
    Wills, EG
    Helenius, A
    Whittaker, GR
    [J]. JOURNAL OF VIROLOGY, 2000, 74 (04) : 1781 - 1786
  • [2] Effect of M1 protein and low pH on nuclear transport of influenza virus ribonucleoproteins
    Bui, M
    Whittaker, G
    Helenius, A
    [J]. JOURNAL OF VIROLOGY, 1996, 70 (12) : 8391 - 8401
  • [3] Several protein regions contribute to determine the nuclear and cytoplasmic localization of the influenza A virus nucleoprotein
    Bullido, R
    Gómez-Puertas, P
    Albo, C
    Portela, A
    [J]. JOURNAL OF GENERAL VIROLOGY, 2000, 81 : 135 - 142
  • [4] Probing single molecules in single living cells
    Byassee, TA
    Chan, WCW
    Nie, SM
    [J]. ANALYTICAL CHEMISTRY, 2000, 72 (22) : 5606 - 5611
  • [5] STRUCTURE OF RIBONUCLEOPROTEIN OF INFLUENZA VIRUS
    COMPANS, RW
    CONTENT, J
    DUESBERG, PH
    [J]. JOURNAL OF VIROLOGY, 1972, 10 (04) : 795 - &
  • [6] MICROINJECTION OF GELSOLIN INTO LIVING CELLS
    COOPER, JA
    BRYAN, J
    SCHWAB, B
    FRIEDEN, C
    LOFTUS, DJ
    ELSON, EL
    [J]. JOURNAL OF CELL BIOLOGY, 1987, 104 (03) : 491 - 501
  • [7] EFFECTS OF CYTOCHALASIN AND PHALLOIDIN ON ACTIN
    COOPER, JA
    [J]. JOURNAL OF CELL BIOLOGY, 1987, 105 (04) : 1473 - 1478
  • [8] IDENTIFICATION OF THE SEQUENCE RESPONSIBLE FOR THE NUCLEAR ACCUMULATION OF THE INFLUENZA-VIRUS NUCLEOPROTEIN IN XENOPUS OOCYTES
    DAVEY, J
    DIMMOCK, NJ
    COLMAN, A
    [J]. CELL, 1985, 40 (03) : 667 - 675
  • [9] DEBRABANDER MJ, 1976, CANCER RES, V36, P905
  • [10] A MONOCLONAL-ANTIBODY AGAINST THE NUCLEAR-PORE COMPLEX INHIBITS NUCLEOCYTOPLASMIC TRANSPORT OF PROTEIN AND RNA INVIVO
    FEATHERSTONE, C
    DARBY, MK
    GERACE, L
    [J]. JOURNAL OF CELL BIOLOGY, 1988, 107 (04) : 1289 - 1297