Difference between revisions of "PMID:1989883"

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!align=left  |Keywords
 
!align=left  |Keywords
 
||
 
||
−
Amino Acid Sequence; Bacterial Proteins; Base Sequence; Chromosomal Proteins, Non-Histone; Chromosomes, Bacterial; Cloning, Molecular; DNA, Bacterial; Escherichia coli; Escherichia coli Proteins; Genes, Bacterial; Genotype; Molecular Sequence Data; Molecular Weight; Mutation; Protein Conformation; Restriction Mapping; Sequence Homology, Nucleic Acid
+
Chromosome; Anucleate cells; Chromosome Partitioning;
  
 
|- class="tableEdit_footer"  
 
|- class="tableEdit_footer"  
Line 34: Line 34:
  
 
==Main Points of the Paper ==
 
==Main Points of the Paper ==
−
*Describes the identification of ''mukB''
+
*''mukB'' gene product is required for chromosome positioning from 22C to 42C
−
*MukB is involved in chromosome partitioning
+
*''mukB'' gene product is not essential at lower temperature, but is essential for cell division at higher temperatures
 +
*''mukB'' is mapped to 21 minutes on the chromosome
 +
*''mukB'' is cloned
 +
* Anucleate cell formation is not due to inhibition of DNA replication
 +
*Nucleotide sequence with putative promoter region and RBS is included
 +
*Predicted amino acid sequence are shown with predicted structural motifs,ATP binding sequence is included
  
 
== Materials and Methods Used ==
 
== Materials and Methods Used ==
−
{{LitMaterials}}
+
*Temperature shifts
 +
*Fluorescence microscopy
 +
*Plating
 +
*Computer analysis of sequence
  
 
==Phenotype Annotations==
 
==Phenotype Annotations==
Line 62: Line 70:
 
''mukB106''
 
''mukB106''
 
|
 
|
−
Aberrant chromosome partitioning at low temperature (conditional)
+
Increased number of anucleate cells at 22C or 30C
 +
 +
 
 +
|
 +
Morphology
 +
|
 +
nucleoid morphology of cell
 +
|
 +
Microscopy
 +
|
 +
Figure 1B
 +
|-
 +
|
 +
''Escherichia coli'' W3110
 +
|
 +
NCBI:316407
 +
|
 +
SH3367
 +
|
 +
''mukB106''
 +
|
 +
Increased number of cell pairs at 22C or 30C
 +
 
  
  
Line 68: Line 98:
 
Morphology
 
Morphology
 
|
 
|
−
nucleoid morphology of cell
+
one anucleate cell with ~1 unit of cell length and one nucleate cell with 1-2 units
 
|
 
|
 
Microscopy
 
Microscopy
 
|
 
|
−
Figure 1
+
Figure 1B, C, D- 
 
|-  
 
|-  
 
|
 
|
Line 83: Line 113:
 
''mukB106''
 
''mukB106''
 
|
 
|
−
Increased number of anucleate cells at low temperature
+
Increased number of cell pairs at 22C or 30C
−
+
 
 +
 
  
 
|
 
|
 
Morphology
 
Morphology
 
|
 
|
−
nucleoid morphology of cell
+
one nucleate cell with 2 copies of chromosomal DNA and one cell having small amounts at the cell pole
 
|
 
|
 
Microscopy
 
Microscopy
 
|
 
|
−
Figure 1
+
Figure 1E, E', F, F'
 +
|-
 +
|
 +
''Escherichia coli'' W3110
 +
|
 +
NCBI:316407
 +
|
 +
GC7528
 +
|
 +
''ΔmukB::kan''
 +
|
 +
Decreased ability to form colonies at high temperature
 +
 
 +
 
 +
 
 +
|
 +
Growth
 +
|
 +
CFUs decreased as the temperature was increased.
 +
|
 +
Plating Assay
 +
|
 +
Figure 4A- tested on a variety of media
 
|-  
 
|-  
 
|
 
|
Line 104: Line 157:
 
''mukB106''
 
''mukB106''
 
|
 
|
−
Increased production of anucleate cells during growth
+
Increased chromosomal guillotining during cell division at 22C or 30C
 +
 
 +
 
 +
 
 +
|
 +
Morphology
 +
|
 +
one nucleate cell with 2 copies of chromosomal DNA and one cell having small amounts at the cell pole
 +
|
 +
Microscopy
 +
|
 +
Figure 1E, E', F, F'
 +
|-
 +
|
 +
''Escherichia coli'' W3110
 +
|
 +
NCBI:316407
 +
|
 +
GC7528
 +
|
 +
''ΔmukB::kan''
 +
|
 +
Increased number of anucleate cells at 22C
 
   
 
   
  
Line 114: Line 189:
 
Microscopy
 
Microscopy
 
|
 
|
−
Table 1
+
Figure 1G
 
|-  
 
|-  
 
|
 
|
Line 123: Line 198:
 
SH3367
 
SH3367
 
|
 
|
−
''ΔmukB''
+
''mukB106''
 
|
 
|
−
Abolished colony forming ability at high temperature
+
Tiny colony formation at 42C
−
+
 
 +
 
  
 
|
 
|
−
Growth
+
Morphology
 
|
 
|
−
plating at high temperature
+
 
 
|
 
|
 
Plating Assay
 
Plating Assay
 
|
 
|
−
Figure 4
+
when grown on L broth overnight
 
|-  
 
|-  
 
|
 
|
Line 146: Line 222:
 
''mukB106''
 
''mukB106''
 
|
 
|
−
Increased number of anucleate cells at low temperature
+
Heterogeneous cell populations at 37C
−
+
 
 +
 
 +
 
 +
|
 +
Morphology
 +
|
 +
Liquid cultures grown at 37C produce nucleate cells of normal cell size, elongated cells with many nucleoids, and anucleated cells
 +
|
 +
Microscopy
 +
|
 +
Figure 1
 +
|-
 +
|
 +
''Escherichia coli'' W3110
 +
|
 +
NCBI:316407
 +
|
 +
GC7528
 +
|
 +
''ΔmukB::kan''
 +
|
 +
Heterogeneous cell populations at 37C
 +
 
 +
 
 +
 
 +
|
 +
Morphology
 +
|
 +
Liquid cultures grown at 37C produce nucleate cells of normal cell size, elongated cells with many nucleoids, and anucleated cells
 +
|
 +
Microscopy
 +
|
 +
Figure 1H
 +
|-
 +
|
 +
''Escherichia coli'' W3110
 +
|
 +
NCBI:316407
 +
|
 +
SH3367
 +
|
 +
''mukB106''
 +
|
 +
Irregular chromosomal distribution within cell at 42C
 +
 
 +
 
  
 
|
 
|
 
Morphology
 
Morphology
 
|
 
|
−
nucleoid morphology of cell
+
 
 
|
 
|
 
Microscopy
 
Microscopy
Line 167: Line 288:
 
''mukB106''
 
''mukB106''
 
|
 
|
−
Partially defective in cell division at high temperature.
+
Elongated cell length at 42C
 +
 
 +
 
 +
 
 
|
 
|
 
Morphology
 
Morphology
 
|
 
|
−
morphology of cell
+
 
 
|
 
|
 
Microscopy
 
Microscopy
Line 188: Line 312:
 
<references/>
 
<references/>
  
−
[[Category:Template]]
+
[[Category:Hiraga's Fluophase Combined Method]]
 +
[[Category:Microscopy, Fluorescence]]
 +
[[Category:SDS-PAGE]]
 +
 
 +
[[Category:Publication]]
 +
[[Category:To Be Converted]]

Latest revision as of 12:09, 22 June 2011

Citation

Niki, H, Jaffé, A, Imamura, R, Ogura, T and Hiraga, S (1991) The new gene mukB codes for a 177 kd protein with coiled-coil domains involved in chromosome partitioning of E. coli.EMBO J. 10:183-93

Abstract

An Escherichia coli temperature sensitive mutant which produces spontaneously normal size anucleate cells at low temperature was isolated. The mutant is defective in a previously undescribed gene, named mukB, located at 21 min on the chromosome. The mukB gene codes for a large protein (approximately 180 kd). A 1534 amino acid protein (176,826 daltons) was deduced from the nucleotide sequence of the mukB gene. Computer analysis revealed that the predicted MukB protein has distinct domains: an amino-terminal globular domain containing a nucleotide binding sequence, a central region containing two alpha-helical coiled-coil domains and one globular domain, and a carboxyl-terminal globular domain which is rich in Cys, Arg and Lys. A 180 kd protein detected in wild-type cell extracts by electrophoresis is absent in mukB null mutants. Although the null mutants are not lethal at low temperature, the absence of MukB leads to aberrant chromosome partitioning. At high temperature the mukB null mutants cannot form colonies and many nucleoids are distributed irregularly along elongated cells. We conclude that the MukB protein is required for chromosome partitioning in E. coli.

Links

PubMed

Keywords

Chromosome; Anucleate cells; Chromosome Partitioning;

edit table

Main Points of the Paper

  • mukB gene product is required for chromosome positioning from 22C to 42C
  • mukB gene product is not essential at lower temperature, but is essential for cell division at higher temperatures
  • mukB is mapped to 21 minutes on the chromosome
  • mukB is cloned
  • Anucleate cell formation is not due to inhibition of DNA replication
  • Nucleotide sequence with putative promoter region and RBS is included
  • Predicted amino acid sequence are shown with predicted structural motifs,ATP binding sequence is included

Materials and Methods Used

  • Temperature shifts
  • Fluorescence microscopy
  • Plating
  • Computer analysis of sequence

Phenotype Annotations

See Help:AnnotationTable for details on how to edit this table.
<protect>

Species Taxon ID Strain Gene (if known) OMP Phenotype Details Evidence Notes

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Increased number of anucleate cells at 22C or 30C


Morphology

nucleoid morphology of cell

Microscopy

Figure 1B

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Increased number of cell pairs at 22C or 30C


Morphology

one anucleate cell with ~1 unit of cell length and one nucleate cell with 1-2 units

Microscopy

Figure 1B, C, D-

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Increased number of cell pairs at 22C or 30C


Morphology

one nucleate cell with 2 copies of chromosomal DNA and one cell having small amounts at the cell pole

Microscopy

Figure 1E, E', F, F'

Escherichia coli W3110

NCBI:316407

GC7528

ΔmukB::kan

Decreased ability to form colonies at high temperature


Growth

CFUs decreased as the temperature was increased.

Plating Assay

Figure 4A- tested on a variety of media

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Increased chromosomal guillotining during cell division at 22C or 30C


Morphology

one nucleate cell with 2 copies of chromosomal DNA and one cell having small amounts at the cell pole

Microscopy

Figure 1E, E', F, F'

Escherichia coli W3110

NCBI:316407

GC7528

ΔmukB::kan

Increased number of anucleate cells at 22C


Morphology

nucleoid morphology of cell

Microscopy

Figure 1G

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Tiny colony formation at 42C


Morphology

Plating Assay

when grown on L broth overnight

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Heterogeneous cell populations at 37C


Morphology

Liquid cultures grown at 37C produce nucleate cells of normal cell size, elongated cells with many nucleoids, and anucleated cells

Microscopy

Figure 1

Escherichia coli W3110

NCBI:316407

GC7528

ΔmukB::kan

Heterogeneous cell populations at 37C


Morphology

Liquid cultures grown at 37C produce nucleate cells of normal cell size, elongated cells with many nucleoids, and anucleated cells

Microscopy

Figure 1H

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Irregular chromosomal distribution within cell at 42C


Morphology

Microscopy

Figure 1

Escherichia coli W3110

NCBI:316407

SH3367

mukB106

Elongated cell length at 42C


Morphology

Microscopy

Figure 1

edit table

</protect>

Notes

References

See Help:References for how to manage references in omp dev.

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