The human myristoylated alanine-rich C kinase substrate (MARCKS) gene (MACS). Analysis of its gene product, promoter, and chromosomal localization.

2.50
Hdl Handle:
http://hdl.handle.net/10541/106558
Title:
The human myristoylated alanine-rich C kinase substrate (MARCKS) gene (MACS). Analysis of its gene product, promoter, and chromosomal localization.
Authors:
Harlan, D M; Graff, J M; Stumpo, D J; Eddy, R L; Shows, T B; Boyle, John M; Blackshear, P J
Abstract:
The expression of a major cellular substrate for protein kinase C, the MARCKS protein, is regulated in a cell-, tissue-, and developmental stage-specific fashion; in addition, this expression can be stimulated acutely by various cytokines in certain cell types. We have begun to characterize the human gene in order to elucidate the genetic elements responsible for this highly regulated expression. We first cloned a human MARCKS cDNA, which encoded a predicted protein of 332 amino acids (Mr 31,600) that was approximately 89, 74, and 59% identical to the bovine, mouse, and chicken proteins, respectively. Regions conserved at the amino acid level included the amino-terminal myristoylation consensus sequence, the site of intron splicing, and the phosphorylation site domain. The human cDNA was used to demonstrate that tumor necrosis factor-alpha could rapidly stimulate MARCKS gene transcription in the human promyelocytic leukemia cell line HL60. Genomic clones were then isolated; sequence analysis identified a putative promoter region that had no TATA box and contained multiple transcription initiation sites in a region spanning 57 base pairs (bp). This was followed by a 5'-untranslated region of approximately 400 bp, which displayed a complex predicted secondary structure with a delta G of -73.4 kcal/mol. Plasmid constructions containing between 52 and 1453 bp of the human MARCKS promoter linked to the human growth hormone gene were then used in transient expression experiments. Constructions containing 52 and 110 bp of the MARCKS promoter did not exhibit promoter function while the larger constructions all exhibited promotor function; the 248-bp fragment of the MARCKS promoter was 80% as effective as the human ferritin promoter in stimulating expression of human growth hormone in intact cells. Using an insert from the human genomic clone as a probe, we identified human chromosome 6, q21-qter, as the location of the MARCKS gene; this has been assigned the gene symbol MACS.
Affiliation:
Howard Hughes Medical Institute Laboratories, Duke University Medical Center, Durham, North Carolina 27710.
Citation:
The human myristoylated alanine-rich C kinase substrate (MARCKS) gene (MACS). Analysis of its gene product, promoter, and chromosomal localization. 1991, 266 (22):14399-405 J. Biol. Chem.
Journal:
Journal of Biological Chemistry
Issue Date:
5-Aug-1991
URI:
http://hdl.handle.net/10541/106558
PubMed ID:
1860846
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorHarlan, D Men
dc.contributor.authorGraff, J Men
dc.contributor.authorStumpo, D Jen
dc.contributor.authorEddy, R Len
dc.contributor.authorShows, T Ben
dc.contributor.authorBoyle, John Men
dc.contributor.authorBlackshear, P Jen
dc.date.accessioned2010-06-21T14:05:40Z-
dc.date.available2010-06-21T14:05:40Z-
dc.date.issued1991-08-05-
dc.identifier.citationThe human myristoylated alanine-rich C kinase substrate (MARCKS) gene (MACS). Analysis of its gene product, promoter, and chromosomal localization. 1991, 266 (22):14399-405 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid1860846-
dc.identifier.urihttp://hdl.handle.net/10541/106558-
dc.description.abstractThe expression of a major cellular substrate for protein kinase C, the MARCKS protein, is regulated in a cell-, tissue-, and developmental stage-specific fashion; in addition, this expression can be stimulated acutely by various cytokines in certain cell types. We have begun to characterize the human gene in order to elucidate the genetic elements responsible for this highly regulated expression. We first cloned a human MARCKS cDNA, which encoded a predicted protein of 332 amino acids (Mr 31,600) that was approximately 89, 74, and 59% identical to the bovine, mouse, and chicken proteins, respectively. Regions conserved at the amino acid level included the amino-terminal myristoylation consensus sequence, the site of intron splicing, and the phosphorylation site domain. The human cDNA was used to demonstrate that tumor necrosis factor-alpha could rapidly stimulate MARCKS gene transcription in the human promyelocytic leukemia cell line HL60. Genomic clones were then isolated; sequence analysis identified a putative promoter region that had no TATA box and contained multiple transcription initiation sites in a region spanning 57 base pairs (bp). This was followed by a 5'-untranslated region of approximately 400 bp, which displayed a complex predicted secondary structure with a delta G of -73.4 kcal/mol. Plasmid constructions containing between 52 and 1453 bp of the human MARCKS promoter linked to the human growth hormone gene were then used in transient expression experiments. Constructions containing 52 and 110 bp of the MARCKS promoter did not exhibit promoter function while the larger constructions all exhibited promotor function; the 248-bp fragment of the MARCKS promoter was 80% as effective as the human ferritin promoter in stimulating expression of human growth hormone in intact cells. Using an insert from the human genomic clone as a probe, we identified human chromosome 6, q21-qter, as the location of the MARCKS gene; this has been assigned the gene symbol MACS.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshBlotting, Northern-
dc.subject.meshCattle-
dc.subject.meshChickens-
dc.subject.meshChromosome Mapping-
dc.subject.meshChromosomes, Human, Pair 6-
dc.subject.meshDNA-
dc.subject.meshHumans-
dc.subject.meshIntracellular Signaling Peptides and Proteins-
dc.subject.meshMembrane Proteins-
dc.subject.meshMice-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshNucleic Acid Conformation-
dc.subject.meshPlasmids-
dc.subject.meshPromoter Regions, Genetic-
dc.subject.meshProtein Kinase C-
dc.subject.meshProteins-
dc.subject.meshRNA, Messenger-
dc.subject.meshSequence Alignment-
dc.subject.meshSequence Homology, Nucleic Acid-
dc.titleThe human myristoylated alanine-rich C kinase substrate (MARCKS) gene (MACS). Analysis of its gene product, promoter, and chromosomal localization.en
dc.typeArticleen
dc.contributor.departmentHoward Hughes Medical Institute Laboratories, Duke University Medical Center, Durham, North Carolina 27710.en
dc.identifier.journalJournal of Biological Chemistryen

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