Fundamental cellular processes do not require vertebrate-specific sequences within the TATA-binding protein

Edward E Schmidt; Alla A Bondareva; Jay R Radke; Mario R Capecchi

doi:10.1074/jbc.M211205200

Back

Fundamental cellular processes do not require vertebrate-specific sequences within the TATA-binding protein

Journal article

Open access

Peer reviewed

Fundamental cellular processes do not require vertebrate-specific sequences within the TATA-binding protein

Edward E Schmidt, Alla A Bondareva, Jay R Radke and Mario R Capecchi

The Journal of biological chemistry, Vol.278(8), pp.6168-6174

02/21/2003

DOI: https://doi.org/10.1074/jbc.M211205200

Handle:

https://hdl.handle.net/2376/107015

PMID: 12471023

Files and links (1)

url

https://doi.org/10.1074/jbc.M211205200View

Published (Version of record) Open

Abstract

Sequence Deletion

Protein Biosynthesis

Species Specificity

Embryo, Mammalian

Fibroblasts - physiology

RNA, Messenger - genetics

Cells, Cultured

Gene Expression Regulation - physiology

DNA Primers

Reverse Transcriptase Polymerase Chain Reaction

Mice, Knockout

TATA-Box Binding Protein - metabolism

Vertebrates

Animals

TATA-Box Binding Protein - genetics

Embryo, Nonmammalian

Transcription, Genetic

Fibroblasts - cytology

Mice

TATA-Box Binding Protein - chemistry

The 180-amino acid core of the TATA-binding protein (TBPcore) is conserved from Archae bacteria to man. Vertebrate TBPs contain, in addition, a large and highly conserved N-terminal region that is not found in other phyla. We have generated a line of mice in which the tbp allele is replaced with a version, tbp(Delta N), which lacks 111 of 135 N-terminal amino acid residues. Most tbp(Delta N/Delta N) fetuses die in midgestation. To test whether a disruption of general cellular processes contributed to this fetal loss, primary fibroblast cultures were established from +/+, Delta N/+, and Delta N/Delta N fetuses. The cultures exhibited no genotype-dependent differences in proliferation or in expression of the proliferative markers dihydrofolate reductase (DHFR) mRNA (S phase-specific) and cdc25B mRNA (G(2)-specific). The mutation had no effect on transcription initiation site fidelity by either RNA polymerase II (pol II) or pol III. Moreover, the mutation did not cause differences in levels of U6 RNA, a pol III-dependent component of the splicing machinery, in mRNA splicing efficiency, in expression of housekeeping genes from either TATA-containing or TATA-less promoters, or in global gene expression. Our results indicated that general eukaryotic cell functions are unaffected by deletion of these vertebrate-specific sequences from TBP. Thus, all activities of this polypeptide domain must either be compensated for by redundant activities or be restricted to situations that are not represented by primary fibroblasts.

Metrics

9 Record Views

Details

Title: Fundamental cellular processes do not require vertebrate-specific sequences within the TATA-binding protein
Creators: Edward E Schmidt - Department of Veterinary Molecular Biology, Marsh Laboratories, Montana State University, Bozeman, Montana 59717, USA. eschmidt@montana.edu
Alla A Bondareva
Jay R Radke
Mario R Capecchi
Publication Details: The Journal of biological chemistry, Vol.278(8), pp.6168-6174
Academic Unit: UNKNOWN
Publisher: United States
Grant note: R03 HD035824-02 / NICHD NIH HHS R03 HD039242-02 / NICHD NIH HHS R03 HD039242-01 / NICHD NIH HHS
Identifiers: 99900546994201842
Language: English
Resource Type: Journal article