Microtubule protein ADP-ribosylation in vitro leads to assembly inhibition and rapid depolymerization

Journal Article · Tue Jan 14 00:00:00 EST 1992 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00116a042· OSTI ID:5560008

Scaife, R M ^[1]; Wilson, L ^[2]; Purich, D L ^[3]

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1. Fred Hutchinson Cancer Research Center, Seattle, WA (United States)
2. Univ. of California, Santa Barbara (United States)
3. Univ. of Florida, Gainesville (United States)

Bovine brain microtubule protein, containing both tubulin and microtubule-associated proteins, undergoes ADP-ribosylation in the presence of ({sup 14}C)NAD{sup +} and a turkey erythrocyte mono-ADP-ribosyltransferase in vitro. The modification reaction could be demonstrated in crude brain tissue extracts where selective ADP-ribosylation of both the {alpha} and {beta} chains of tubulin and of the high molecular weight microtubule-associated protein MAP-2 occurred. In experiments with purified microtubule protein, tubulin dimer, the high molecular weight microtubule-associated protein MAP-2, and another high molecular weight microtubule-associated protein which may be a MAP-1 species were heavily labeled. Tubulin and MAP-2 incorporated ({sup 14}C)ADP-ribose to an average extent of approximately 2.4 and 30 mol of ADP-ribose/mol of protein, respectively. Assembly of microtubule protein into microtubules in vitro was inhibited by ADP-ribosylation, and incubation of assembled steady-state microtubules with ADP-ribosyltransferase and NAD{sup +} resulted in rapid depolymerization of the microtubules. Thus, the eukaryotic enzyme can ADP-ribosylate tubulin and microtubule-associated proteins to much greater extents than previously observed with cholera and pertussis toxins, and the modification can significantly modulate microtubule assembly and disassembly.

• OSTI ID: 5560008
• Journal Information: Biochemistry; (United States), Vol. 31:1; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CARBON 14 COMPOUNDS
UPTAKE
MICROTUBULES
POST-TRANSLATION MODIFICATION
ADP
BRAIN
CATTLE
ELECTROPHORESIS
ERYTHROCYTES
TRANSFERASES
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CARBON COMPOUNDS
CELL CONSTITUENTS
CENTRAL NERVOUS SYSTEM
DOMESTIC ANIMALS
ENZYMES
LABELLED COMPOUNDS
MAMMALS
MATERIALS
NERVOUS SYSTEM
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RUMINANTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques