Comprehensive analysis of the Arabidopsis genome revealed a total of 13 sHsps belonging to 6 classes defined on the basis of their intracellular localization and sequence relatedness plus 6 ORFs encoding proteins distantly related to the cytosolic class Cl or the plastidial class of sHsps. The complexity of the Arabidopsis sHsp family far exceeds that in any other organism investigated to date. Furthermore, we have identified a new family of ORFs encoding multidomain proteins that contain one or more regions with homology to the ACD (Acd proteins). The functions of the Acd proteins and the role of their ACDs remain to be investigated.

The small heat shock proteins (sHsps

Small heat shock proteins (sHSPs) are virtually ubiquitous stress proteins that are also found in many normal tissues and accumulate in diseases of protein folding. They generally act as ATP-independent chaperones to bind and stabilize denaturing proteins that can be later reactivated by ATP-dependent Hsp70/DnaK, but the mechanism of substrate capture by sHSPs remains poorly understood. A majority of sHSPs form large oligomers, a property that has been linked to their effective chaperone action. We describe AtHsp18.5 from Arabidopsis thaliana, demonstrating it is dimeric and exhibits robust chaperone activity, adding support to the model that suboligomeric sHSP forms are a substrate binding species. Notably, like oligomeric sHSPs, when bound to substrate AtHsp18.5 assembles into large complexes, indicating reformation of sHSP oligomeric contacts are not required for assembly of sHSP-substrate complexes. Monomers of AtHsp18.5 freely exchange between dimers, but fail to coassemble in vitro with dodecameric plant cytosolic sHSPs, suggesting AtHsp18.5 does not interact by coassembly with these other sHSPs in vivo. Data from controlled proteolysis and hydrogen-deuterium exchange coupled with mass spectrometry show that the N- and C-termini of AtHsp18.5 are highly accessible and lack stable secondary structure, most likely a requirement for substrate interaction. Chaperone activity of a series of AtHsp18.5 truncation mutants confirm that the N-terminal arm is required for substrate protection and that different substrates interact differently with the N-terminal arm. In total, these data imply that the core α-crystallin domain of the sHSPs is a platform for flexible arms that capture substrates to maintain their solubility.

We describe a new class of plant small heat stress proteins (sHsps) with dominant nuclear localization (Hsp17-CIII). The corresponding proteins in tomato, Arabidopsis, and rice are encoded by unique genes containing a short intron in the beta4-encoding region of the alpha-crystallin domain (ACD). The strong nuclear localization results from a cluster of basic amino acid residues in the loop between beta5 and beta6 of the ACD. Using yeast 2-hybrid tests, analyses of native complexes of the sHsps, and immunofluorescence data, we demonstrate that, in contrast to earlier observations (Kirschner et al 2000), proteins of the sHsp classes CI, CII, and CIII interact with each other, thereby influencing oligomerization state and intracellular localization.

The temporal profiles of the emission from titania particle suspensions in Rhodamine 640 perchlorate dye solutions excited by 10-ps pulses of 527-nm radiation were measured over a wide range of particle and dye concentrations and laser powers. The dynamics of stimulated emission from random media is modeled by a random walk of photons within the colloid and rate equations for molecular excitations. The pulse width and the dependence of the threshold for laser action on dye and scatter concentration are computed by a Monte Carlo simulation of the model and are found to be in qualitative agreement with experiment.