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The Resource Molecular genetic analysis of nucleotide excision repair genes in Dictyostelium discoideum, by Sung-Keun Lee

Molecular genetic analysis of nucleotide excision repair genes in Dictyostelium discoideum, by Sung-Keun Lee

Creator
Summary
The DNA helicases encoded by the Xeroderma pigmentosum (XP) B and D genes function in both nucleotide excision repair (NER) and transcription initiation. XPB and XPD patients can exhibit neurological and developmental abnormalities. To better understand the developmental roles of these genes, we used the cellular slime mold Dictyostelium discoideum as a model system. Although Dictyostelium discoideum has only two cell types, stalk and spore cells, its development exhibits many facets of cell differentiation and morphogenesis seen in higher eucaryotes. We have identified and characterized the repB and repD genes from Dictyostelium discoideum. Both genes encode DNA helicases of the SF2 superfamily of helicases. The repD gene contains no introns and the repB gene contains only one intron, which makes their genomic structures dramatically different from the corresponding genes in mammals and fish. However the predicted Dictyostelium proteins share high homology with the human XPB and XPD proteins. The single copy of the repB and D genes map to chromosomes 3 and 1, respectively. The expression of repB and D (and the previously isolated repE) genes during multicellular development was examined, and it was determined that each rep gene has a unique pattern of expression. The pattern and extent of expression of these genes was not affected by the growth history of the cells. Homologous recombination was used to construct C-terminal repB deletion mutants which resembled the mutations found in XPB patients who showed UV-sensitivity and developmental abnormalities. The C-terminal deletion mutants confer UV-sensitivity but did not have any gross developmental phenotype. This result suggests that a second mutant gene may be responsible for the developmental abnormalities in XPB patients. In addition, we have characterized the mntA gene from Dictyostelium discoideum which encodes the $\beta$-1,4-mannosyltransferase enzyme. The mntA gene contains a single small intron and encodes a 493 amino acid protein with a predicted molecular size of 56 kDa. It is located 5$\sp\prime$ to the repE gene on chromosome IV and is transcribed in the opposite orientation to repE with which it shares a 585 base pair of upstream intergenic region. The mntA gene product shares 38% homology with the S. cerevisiae ALG1 gene product. Northern analysis revealed that the expression of the mntA gene is regulated during multicellular development with two periods of mRNA accumulation. The mntA gene is the first Dictyostelium homolog of a gene encoding a protein that is active in the endoplasmic reticulum. (Abstract shortened by UMI.)
Language
eng
Work
Publication
Extent
xiv, 146 leaves
Note
  • Typescript
  • Vita
Instance
Label
Molecular genetic analysis of nucleotide excision repair genes in Dictyostelium discoideum
Title
Molecular genetic analysis of nucleotide excision repair genes in Dictyostelium discoideum
Statement of responsibility
by Sung-Keun Lee
Title variation
Nucleotide excision repair genes in Dictyostelium
Creator
Subject
Language
eng
Summary
The DNA helicases encoded by the Xeroderma pigmentosum (XP) B and D genes function in both nucleotide excision repair (NER) and transcription initiation. XPB and XPD patients can exhibit neurological and developmental abnormalities. To better understand the developmental roles of these genes, we used the cellular slime mold Dictyostelium discoideum as a model system. Although Dictyostelium discoideum has only two cell types, stalk and spore cells, its development exhibits many facets of cell differentiation and morphogenesis seen in higher eucaryotes. We have identified and characterized the repB and repD genes from Dictyostelium discoideum. Both genes encode DNA helicases of the SF2 superfamily of helicases. The repD gene contains no introns and the repB gene contains only one intron, which makes their genomic structures dramatically different from the corresponding genes in mammals and fish. However the predicted Dictyostelium proteins share high homology with the human XPB and XPD proteins. The single copy of the repB and D genes map to chromosomes 3 and 1, respectively. The expression of repB and D (and the previously isolated repE) genes during multicellular development was examined, and it was determined that each rep gene has a unique pattern of expression. The pattern and extent of expression of these genes was not affected by the growth history of the cells. Homologous recombination was used to construct C-terminal repB deletion mutants which resembled the mutations found in XPB patients who showed UV-sensitivity and developmental abnormalities. The C-terminal deletion mutants confer UV-sensitivity but did not have any gross developmental phenotype. This result suggests that a second mutant gene may be responsible for the developmental abnormalities in XPB patients. In addition, we have characterized the mntA gene from Dictyostelium discoideum which encodes the $\beta$-1,4-mannosyltransferase enzyme. The mntA gene contains a single small intron and encodes a 493 amino acid protein with a predicted molecular size of 56 kDa. It is located 5$\sp\prime$ to the repE gene on chromosome IV and is transcribed in the opposite orientation to repE with which it shares a 585 base pair of upstream intergenic region. The mntA gene product shares 38% homology with the S. cerevisiae ALG1 gene product. Northern analysis revealed that the expression of the mntA gene is regulated during multicellular development with two periods of mRNA accumulation. The mntA gene is the first Dictyostelium homolog of a gene encoding a protein that is active in the endoplasmic reticulum. (Abstract shortened by UMI.)
Additional physical form
Also available on the Internet.
Cataloging source
MUU
http://library.link/vocab/creatorDate
1960-
http://library.link/vocab/creatorName
Lee, Sungkeun
Degree
Ph. D.
Dissertation year
1997.
Government publication
government publication of a state province territory dependency etc
Granting institution
University of Missouri-Columbia
Illustrations
illustrations
Index
no index present
Literary form
non fiction
Nature of contents
bibliography
http://library.link/vocab/subjectName
  • Dictyostelium discoideum
  • Molecular genetics
  • Nucleotides
  • Plant genetics
  • DNA repair
  • DNA replication
  • Genetic transcription
  • Genetic recombination
  • Introns
Target audience
specialized
Label
Molecular genetic analysis of nucleotide excision repair genes in Dictyostelium discoideum, by Sung-Keun Lee
Instantiates
Publication
Note
  • Typescript
  • Vita
Bibliography note
Includes bibliographical references (leaves 124-125)
Carrier category
online resource
Carrier category code
cr
Carrier MARC source
rdacarrier
Content category
text
Content type code
txt
Content type MARC source
rdacontent
Control code
42190946
Dimensions
29 cm
Dimensions
unknown
Extent
xiv, 146 leaves
Media category
computer
Media MARC source
rdamedia
Media type code
c
Other physical details
illustrations
Specific material designation
remote
Label
Molecular genetic analysis of nucleotide excision repair genes in Dictyostelium discoideum, by Sung-Keun Lee
Publication
Note
  • Typescript
  • Vita
Bibliography note
Includes bibliographical references (leaves 124-125)
Carrier category
online resource
Carrier category code
cr
Carrier MARC source
rdacarrier
Content category
text
Content type code
txt
Content type MARC source
rdacontent
Control code
42190946
Dimensions
29 cm
Dimensions
unknown
Extent
xiv, 146 leaves
Media category
computer
Media MARC source
rdamedia
Media type code
c
Other physical details
illustrations
Specific material designation
remote

Subject

Genre

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