Biomedical subjects
M J Ackermann
Publications and source records attributed to M J Ackermann.
p34cdc2-mediated phosphorylation at T124 inhibits nuclear import of SV-40 T antigen proteins.
The nuclear import of transcription regulatory proteins appears to be used by the cell to trigger transitions in cell cycle, morphogenesis, and transformation. We have previously observed that the rate at which SV-40 T antigen fusion proteins containing a functional nuclear localization sequence (NLS; residues 126-132) are imported into the nucleus is enhanced in the presence of the casein kinase II (CK-II) site S111/112. In this study purified p34cdc2 kinase was used to phosphorylate T antigen proteins specifically at T124 and kinetic measurements at the single-cell level performed to assess its effect on nuclear protein import. T124 phosphorylation, which could be functionally simulated by a T-to-D124 substitution, was found to reduce the maximal extent of nuclear accumulation whilst negligibly affecting the import rate. The inhibition of nuclear import depended on the stoichiometry of phosphorylation. T124 and S111/112 could be phosphorylated independently of one another. Two alternative mechanisms were considered to explain the inhibition of nuclear import by T124 phosphorylation: inactivation of the NLS and cytoplasmic retention, respectively. Furthermore, we speculate that in vivo T124 phosphorylation may regulate the small but functionally significant amount of cytoplasmic SV-40 T antigen. A sequence comparison showed that many transcription regulatory proteins contain domains comprising potential CK-II-sites, cdc2-sites, and NLS. This raises the possibility that the three elements represent a functional unit regulating nuclear protein import.
Quantification of high pressure nervous syndrome (HPNS) tremor in the guinea pig.
Previous studies have demonstrated two tremorgenic systems that involve separate brain mechanisms and exhibit different peak frequencies. One system (the thalamo-cortical) generates low frequency (4--8 Hz) tremor; the other (the olivo-cerebellar) produces high frequency (10--18 Hz) tremor. Based on this evidence, the present study focused on determining whether one or both of these tremor systems is involved in the high pressure nervous syndrome (HPNS). Specifically, the concern was to identify and to quantify amplitude and frequency characteristics of HPNS tremor in 8 guinea pigs breathing helium-oxygen during compression (40 ft/min) in a chamber dive to 61.6 ATA (2000 fsw) with a bottom time of 1 h. Rectal temperature was recorded and maintained at 39 degrees C +/- 1 degree. Leg tremor was recorded by magnetic inductance and stored on magnetic tape for power spectral analysis. Frequency histograms of the tremor data revealed development of a biphasic response. From surface to about 31.3 ATA (1000 fsw), a low-power, single, 4- to 6-Hz component was evident, which resembled fine or moderate tremor. Between 34.3 ATA (1100 fsw) and 61.6 ATA, a 12- to 18-Hz component emerged abruptly with a dramatic increase in power, which reflected coarse, uncontrollable tremors. In the first 5 to 10 min after the animals arrived at maximum pressure, relative power of the high frequency component dropped to and remained near base-line levels. These results support the hypothesis that HPNS tremor consists of two components and possibly two separate tremor systems.