VISUALIZATION OF THE SUBCELLULAR LOCATION OF SPORULATION PROTEINS IN BACILLUS-SUBTILIS USING IMMUNOFLUORESCENCE MICROSCOPY

被引:124
作者
POGLIANO, K
HARRY, E
LOSICK, R
机构
[1] Dept. of Molec. and Cellular Biology, Biological Laboratories, Harvard University, Cambridge
[2] Department of Biochemistry, University of Sydney, Sydney
关键词
D O I
10.1111/j.1365-2958.1995.mmi_18030459.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We describe the application of immunofluorescence microscopy to visualization of the subcellular localization of proteins involved in coat morphogenesis and chromosome packaging during the process of sporulation in Bacillus subtilis. In confirmation and extension of previous findings, we show that SpolVA, which is responsible for guiding coat formation to the surface of the outer membrane that surrounds the developing spore, assembles into a shell that is located close to or on the surface of this enveloping membrane. CotE, which is responsible for the formation of the outer layer of the coat, assembles into a second shell of apparently larger diameter, Assembly of SpolVA could be detected as early as the morphological stage of polar septation and closely followed the enveloping membrane of the mother cell during the stage of engulfment, thereby providing a sensitive and diagnostic marker for this phagocytic-like process. Surprisingly, the chromosome of the developing spore and the small, acid-soluble proteins, known as alpha/beta-type SASPs, that are known to coat the spore chromosome, were found to co-localize to a doughnut-like ring of approximately 1 mu m in diameter. The use of a double mutant lacking the alpha/beta-type SASP demonstrated that these high abundance, DNA-binding proteins are responsible for packaging the chromosome of the developing spore into this unusual structure, We conclude that sporulation in B. subtilis is a fertile system for addressing cell biological problems in a bacterium and that immunofluorescence microscopy provides a sensitive method for visualizing protein subcellular localization at high resolution.
引用
收藏
页码:459 / 470
页数:12
相关论文
共 42 条
[1]  
ANDERSON W, 1969, J CELL SCI, V4, P611
[2]   FTSZ RING STRUCTURE ASSOCIATED WITH DIVISION IN ESCHERICHIA-COLI [J].
BI, E ;
LUTKENHAUS, J .
NATURE, 1991, 354 (6349) :161-164
[3]   GENE-EXPRESSION IN SINGLE CELLS OF BACILLUS-SUBTILIS - EVIDENCE THAT A THRESHOLD MECHANISM CONTROLS THE INITIATION OF SPORULATION [J].
CHUNG, JD ;
STEPHANOPOULOS, G ;
IRETON, K ;
GROSSMAN, AD .
JOURNAL OF BACTERIOLOGY, 1994, 176 (07) :1977-1984
[4]   THE ACTIN-BASED MOTILITY OF THE FACULTATIVE INTRACELLULAR PATHOGEN LISTERIA-MONOCYTOGENES [J].
COSSART, P ;
KOCKS, C .
MOLECULAR MICROBIOLOGY, 1994, 13 (03) :395-402
[5]   SUBCELLULAR-LOCALIZATION OF PROTEINS INVOLVED IN THE ASSEMBLY OF THE SPORE COAT OF BACILLUS-SUBTILIS [J].
DRIKS, A ;
ROELS, S ;
BEALL, B ;
MORAN, CP ;
LOSICK, R .
GENES & DEVELOPMENT, 1994, 8 (02) :234-244
[6]  
EIKBUSH TH, 1978, CELL, V13, P295
[7]  
ERRINGTON J, 1993, MICROBIOL REV, V57, P1
[8]   INSITU HYBRIDIZATION TO THE CRITHIDIA-FASCICULATA KINETOPLAST REVEALS 2 ANTIPODAL SITES INVOLVED IN KINETOPLAST DNA-REPLICATION [J].
FERGUSON, M ;
TORRI, AF ;
WARD, DC ;
ENGLUND, PT .
CELL, 1992, 70 (04) :621-629
[9]   KINETOPLAST DNA-REPLICATION - MECHANISTIC DIFFERENCES BETWEEN TRYPANOSOMA-BRUCEI AND CRITHIDIA-FASCICULATA [J].
FERGUSON, ML ;
TORRI, AF ;
PEREZMORGA, D ;
WARD, DC ;
ENGLUND, PT .
JOURNAL OF CELL BIOLOGY, 1994, 126 (03) :631-639
[10]   IMMUNOELECTRON MICROSCOPIC LOCALIZATION OF SMALL, ACID-SOLUBLE SPORE PROTEINS IN SPORULATING CELLS OF BACILLUS-SUBTILIS [J].
FRANCESCONI, SC ;
MACALISTER, TJ ;
SETLOW, B ;
SETLOW, P .
JOURNAL OF BACTERIOLOGY, 1988, 170 (12) :5963-5967