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Proteolytic processing of phage lambda tail protein gpH: timing of the cleavage.

We describe a method for the rapid partial purification of intermediate structures of phage lambda tail assembly, using formaldehyde-fixed Escherichia coli cells to precipitate tail-related structures. The purification depends on the specific interaction between the E. coli lambda receptor protein and lambda tail protein gpJ. Protein compositions of tail assembly intermediates were analyzed to determine when in the assembly sequence the minor tail protein gpH is cleaved. gpH joins the tail precursor structure early in the pathway, during assembly of the initiator (a structure that becomes the tail tip). However, gpH is not cleaved until after initiator assembly is complete and after the tail shaft has polymerized onto the initiator. These results suggest that each gpH molecule is extended along the length of the tail. Our results also appear to eliminate an ambiguity in the tail assembly pathway determined by earlier experiments: we argue that gene G acts between genes H and M.

Bacterial Outer Membrane Proteins↗

Intersegmental neuronal pathways in sacrococcygeal spinal cord (S3-Co3) activated by electrical stimulation of tail muscle nerves with low threshold in low spinal cats.

The aim of this study was to elucidate the neuronal pathways mediating propriospinal reflexes of the tail with special reference to the intersegmental connections between group I muscle afferents and motoneurons. Experiments were performed on 25 unanesthetized, spinalized (L1) cats of both sexes. The effects of electrical stimulation of the afferents embedded in different levels of the following muscles: m. extensor caudae medialis (ECM), m. extensor caudae lateralis (ECL), m. flexor caudae longus (FCL), and m. flexor caudae brevis (FCB), were observed in tail motoneurons in the first or second coccygeal (Co1 or Co2) spinal segment. Stimulation of group I muscle afferents from the cranial parts of the tail muscles produced predominantly excitatory postsynaptic potentials. In ECM and FCB, some motoneurons had significant excitatory effects exerted by group I afferent inputs from homonymous and/or synergistic muscles during vertical tail movement, and some motoneurons had predominately excitatory effects from antagonistic muscles during vertical tail movement. Excitatory effects in ECL and FCL motoneurons occurred more often and were larger in size after stimulating the ipsilateral tail muscles than after stimulating the contralateral tail muscles. The stimulation of group I muscle afferents from the caudal parts of the tail muscles produced predominantly inhibitory effects. The shortest neuronal pathways for excitatory postsynaptic potentials and inhibitory postsynaptic potentials were mono- and disynaptic neuronal pathways, respectively. The results of the present experiments suggest that continuous tail movements along the tail are performed by group I reflex pathways.

Afferent Pathways↗

Tail biting in pigs.

One of the costly and welfare-reducing problems in modern pig production is tail biting. Tail biting is an abnormal behaviour, characterized by one pig's dental manipulation of another pig's tail. Tail biting can be classified into two groups: the pre-injury stage, before any wound on the tail is present, and the injury stage, where the tail is wounded and bleeding. Tail biting in the injury stage will reduce welfare of the bitten pig and the possible spread of infection is a health as well as welfare problem. The pigs that become tail biters may also suffer, because they are frustrated due to living in a stressful environment. This frustration may result in an excessive motivation for biting the tails of pen mates. This review aims to summarize recent research and theories in relation to tail biting.

Animal Welfare↗

Neural circuit of tail-elicited siphon withdrawal in Aplysia. II. Role of gated inhibition in differential lateralization of sensitization and dishabituation.

In the preceding report, we observed that tail-shock-induced sensitization of tail-elicited siphon withdrawal reflex (TSW) of Aplysia was expressed ipsilaterally but that dishabituation induced by an identical tail shock was expressed bilaterally. Here we examined the mechanisms of this differential lateralization. We first isolated the modulatory pathway responsible for the induction of contralateral dishabituation by making selective nerve cuts. We found that an intact pleural-abdominal connective, the descending pathway connecting the ring ganglia with the abdominal ganglion, ipsilateral to the shock was required for contralateral dishabituation. We examined whether network inhibition suppresses the contralateral effects of tail shock in nonhabituated preparations. We found that blockade of inhibitory transmission in the CNS by the nicotinic ACh inhibitor d-tubocurarine (d-TC) rendered tail shock capable of inducing bilateral sensitization. We next asked whether serotonin (5-HT), a neuromodulator released in the CNS in response to tail shock, was affected by d-TC. We found that d-TC does not alter 5-HT processes in the ring ganglia: it had no effect on the lateralized pattern of tail nerve shock-induced changes in tail sensory neuron excitability, a 5-HT-dependent process, and it did not alter tail nerve shock-evoked release of 5-HT. By contrast, d-TC enhanced 5-HT release in the abdominal ganglion. Consistent with this observation, restricting d-TC to the abdominal ganglion rendered tail nerve shock capable of producing bilateral sensitization. Together with the results of the preceding paper, our results suggest a model in which TSW sensitization and dishabituation can be dissociated both anatomically and mechanistically.

Animals↗

Midbrain suppression of limb withdrawal and tail flick reflexes in the rat: correlates with descending inhibition of sacral spinal neurons.

1. The aim of this study was to utilize new quantitative behavioral methods in rats to investigate the effects of electrical stimulation in midbrain analgesia areas on the magnitude of flexion hindlimb withdrawal and tail flick reflexes evoked by graded noxious heating. Electrophysiological experiments were then done with the use of these animals to correlate behavioral data with the effects of identical midbrain stimulation on sacral dorsal horn neuronal responses to graded heating of the tail. 2. To quantify limb withdrawals, electromyographs (EMGs) were recorded in biceps femoris during withdrawals elicited by noxious heat stimuli (40-52 degrees C, 5 s, 2-min intervals) delivered to the plantar surface of the hind paw, without and during concomitant electrical stimulation (100 ms, 100-Hz trains, 3/s, 10-600 microA) in midbrain periaqueductal gray (PAG) or laterally adjacent reticular formation (LRF) via previously implanted electrodes. The same animals were tested with the use of a tail flick paradigm modified to allow measurement of the force of tail movements in three orthogonal planes and thereby calculate the overall force vector of tail flicks elicited by graded noxious radiant heat pulses (38-58 degrees C, 5 s, 2-min intervals) delivered to the tail, again with and without concomitant PAG or LRF stimulation. Finally, the same rats were anesthetized with pentobarbital sodium and microelectrode recordings made from single sacral dorsal horn neurons responsive to the same noxious heat stimuli delivered to the tail to assess effects of PAG and LRF stimulation. 3. PAG and LRF stimulation suppressed the magnitude of limb flexor EMGs, and tail flick force vectors, in an intensity-dependent manner. Recruitment of suppression of both limb withdrawal EMGs and tail flicks was generally more effective for LRF compared with PAG stimulation, although mean thresholds for suppression were similar. Tail flick force and limb withdrawal EMGs recorded from the same rat in separate sessions were suppressed about equally in a majority of cases. 4. Limb withdrawal EMG magnitude increased monotonically from threshold (approximately 40 degrees C) to 52 degrees C. The population stimulus-response function was fit equally well by linear regression or a 2 degrees polynomial function (r2 = 0.79 for both). PAG stimulation significantly reduced the mean slope of the stimulus-response function (to 73%; n = 15), whereas LRF stimulation shifted it toward the right with a smaller slope reduction (to 85%) and 3 degrees C increase in threshold.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Short communication: behavioral and physiological indicators of sensitivity or chronic pain following tail docking.

Docking the tails of dairy cattle causes mild to moderate behavior changes and physiological indicators of acute pain, but no studies have investigated the possibility that tail docking may lead to chronic pain. In human amputees, an incidence of increased limb surface temperature is associated with phantom limb pain, a central nervous system representation that survives peripheral loss. The objectives of this study were to assess indicators of sensitivity or chronic pain in heifers by using behavioral indicators and thermography. We tested 14 Holstein heifers, 7 docked and 7 intact, from a previous neonatal tail-docking experiment. All 14 animals were videotaped during a test sequence of alternating cold (-9 degrees C), hot (54 degrees C), and neutral packs applied to the underside of the tail. Packs were placed approximately 30.5 cm from the tail head on all animals. A thermal image of the tail was taken using infrared imagery prior to and after temperature sensitivity testing. Docked heifers tended to have greater changes in surface temperatures following the test sequence than did nondocked heifers. In docked heifers, temperatures on the underside of the tail were higher than those at the tip of the tail, both prior to and following the test sequence. Docked heifers also showed substantially higher stomping activity following application of the cold pack. Shifting increased in intact heifers after application of the hot pack, but shifting of the docked heifers did not change. Greater changes were observed in the tail surface temperatures of the docked heifers following temperature manipulation, similar to human amputees who are experiencing phantom limb pain, indicating that similar mechanisms are present in the stump of the docked tail. The behaviors of docked heifers indicated changes in their sensitivity to heat and cold.

Animal Welfare↗

The influence of tail biting on performance of fattening pigs.

In comparison to 29 non bitten animals, severe tail biting was found to decrease the daily weight gain (DWG) by 25% in 8 fattening pigs during the period of biting. However, when comparing the weight gain of the lifetime between bitten and non bitten pigs, no influence of the tail biting was found. It is of interest that severely wounded pigs were parenterally treated with prokainpenicillin G for 3 consecutive days in connection with the tail biting, which could be suggested to promote the growth by reducing the influence of infections gained by the tail biting as well as of other infections present in herds rearing conventional pigs. Despite penicillin treatment, abscesses were more frequently recorded in tail bitten pigs than in non bitten animals. The tail biting was not equally distributed between the sexes, as barrows were more frequently bitten than gilts. Among the unbitten pigs, barrows were also found to grow faster than gilts. Indeed, when comparing tail bitten and non bitten barrows, a negative influence of tail biting on DWG was not only shown during the period of biting, but could also be monitored as a reduced DWG from that period until slaughter by 11% and during lifetime by 5% (the tail bitten gilts were too few to allow statistical calculations). These results clearly indicate that tail biting affects the growth rate of the lifetime despite penicillin treatment. However, it should be stressed that this decreased lifetime DWG may not be monitored when evaluating abattoir data because the sex distribution of the pigs may not be known in such materials.

Abscess↗

Evidence of a role for cell death in the disappearance of the embryonic human tail.

The development and disappearance of the human tail between stages 14 and 22 were studied using scanning and transmission electron microscopy, supravital staining and light microscopy. The tail is a prominent feature of the human embryo during stage 14 and is composed of paired somites, mesenchyme and extensions of the neural tube, notochord and gut. The tail grows with the embryo through early stage 17 when it extends more than a millimeter from the trunk. Overgrowth by the trunk at the base of the tail may account for the loss of part of its length during late stage 17 and stage 18. However, during stage 17 cells begin to die in all structures throughout the tail. Cell death continues in the succeeding stages reaching massive numbers by stages 18 and 19, and the tail becomes less and less prominent with developmental time. Most of the dead cells are phagocytosed. The debris-laden macrophages appear to migrate from the tail to the body. By late stage 21 or early stage 22 there is no free tail. We conclude that cell death has a major role in the destruction of tail structures and the concurrent loss of the human tail.

Cell Survival↗

Intact alpha-actinin molecules are needed for both the assembly of actin into the tails and the locomotion of Listeria monocytogenes inside infected cells.

After the infectious bacterium, Listeria monocytogenes, is phagocytosed by a host cell, it leaves the lysosome and recruits the host cell's cytoskeletal proteins to assemble a stationary tail composed primarily of actin filaments cross-linked with alpha-actinin. The continual recruitment of contractile proteins to the interface between the bacterium and the tail accompanies the propulsion of the bacterium ahead of the elongating tail. When a bacterium contacts the host cell membrane, it pushes out the membrane into an undulating tubular structure or filopodium that envelops the bacterium at the tip with the tail of cytoskeletal proteins behind it. Previous work has demonstrated that alpha-actinin can be cleaved into two proteolytic fragments whose microinjection into cells interferes with stress fiber integrity. Microinjection of the 53 kD alpha-actinin fragment into cells infected with Listeria monocytogenes, induces the loss of tails from bacteria and causes the bacteria to become stationary. Infected cells that possess filopodia when injected with the 53 kD fragment lose their filopodia. These results indicate that intact alpha-actinin molecules play an important role in the intracellular motility of Listeria, presumably by stabilizing the actin fibers in the stationary tails that are required for the bacteria to move forward. Fluorescently labeled vinculin associated with the tails when it was injected into infected cells. Talin antibody staining indicated that this protein, also, is present in the tails. These observations suggest that the tails share properties of attachment plaques normally present in the host cells. This model would explain the ability of the bacterium (1) to move within the cytoplasm and (2) to push out the surface of the cell to form a filopodium. The attachment plaque proteins, alpha-actinin, talin, and vinculin, may bind and stabilize the actin filaments as they polymerize behind the bacteria and additionally could also enable the tails to bind to the cell membrane in the filopodia.

Actinin↗

Effects of tail-like substituents on the binding of competitive inhibitors to the Q(B) site of photosystem II.

The QB quinone-binding site of photosystem II is an important target for herbicides. Two major classes of herbicides are based on s-triazine and phenylurea moieties. A small library of triazine and phenylurea compounds has been synthesized which have tail-like substituents in order to test the effects of charge, hydrophobicity and size of the tail on binding properties. It is found that a tail can be attached to one of the alkylamino groups of triazine-type herbicides or to the para position of phenylurea-type herbicides without loss of binding, provided that the tail is hydrophobic. This indicates that the herbicides must be oriented in the QB site such that these positions point toward the natural isoprenyl tail-binding pocket that extends out of the Q(B) site. In turn, the requirement that the tail must extend out of the QB site constrains the size of the other herbicide substituents in the pocket. This is in agreement with the presumed orientation and fit of ligands in the QB site. When longer hydrophobic tails are used, the binding penalty that occurs upon adding a charged substituent at the distal end is reduced. This allows the use of a series of tail substituents possessing a distal charge as an approximate molecular ruler to measure the distance from the QB site to the aqueous phase. Even a 10-carbon alkyl chain still shows a 4-fold effect from the presence or absence of a distal charge. Such a chain does not appear to be long enough to extend from the bulk aqueous phase to the QB site because binding is completely lost when a large hydrophilic domain (PEG(4000)) is attached to the distal end. Longer tails are effective only if they are sufficiently hydrophobic. An effort was made to use tailed herbicides for affinity binding of photosystem II. It was found that hydrophobic linkers promote nonspecific binding, but careful choice of solvent conditions, such as the use of excess nonionic detergent well above its critical micelle concentration, might obviate this problem during affinity-binding applications.

Binding, Competitive↗

Sexual selection for white tail spots in the barn swallow in relation to habitat choice by feather lice.

Many bird species have white spots in their tails or wing feathers, and such characters have been hypothesized to be either reliable signals (handicaps) or amplifiers that facilitate the message of a signal. In barn swallows, Hirundo rustica, the size of the white spots in the tail feathers is sexually dimorphic and positively correlated with feather length. We tested whether such spots act as handicaps or amplifiers. These white spots affect sexual selection in barn swallows, as shown by an experiment in which we randomly subjected males to (1) a considerable reduction of the size of all the spots by the use of a black permanent marker pen, (2) a small reduction of the size of the spots, or (3) no reduction. There was a positive association between spot size and the number of offspring produced per season. The white tail spots were preferred by feather-eating Mallophaga as a feeding site: holes made by Mallophaga were more abundant in the white spots than expected by chance. A habitat choice experiment with Mallophaga on barn swallow tail feathers revealed that they preferred white spots over black parts of the tail feathers. We therefore expected long-tailed male barn swallows to have more Mallophaga than short-tailed males. However, the opposite relationship was observed, indicating that long-tailed males may reliably signal their quality by the presence of large white tail spots without parasite damage. Thus white tail spots in barn swallows appear to be a reliable signal of phenotypic quality. Copyright 1999 The Association for the Study of Animal Behaviour.

Journal Article↗

Nucleosome dynamics. VI. Histone tail regulation of tetrasome chiral transition. A relaxation study of tetrasomes on DNA minicircles.

We have recently described the relaxation of mononucleosomes on an homologous series of 351-366 bp DNA minicircles, as a tool to study nucleosome structure and dynamics in vitro. Nucleosomes were found to have a tail-regulated access to three distinct DNA conformations, depending on the crossing between the entering and exiting DNAs, and its polarity. This approach was now used to explore tetrasome chiral transition, and the influence of the histone tails. The data confirmed the existence of two states, with linking number differences DeltaLk(t)=-0.74(+/-0.01) and +0.51(+/-0.06). As expected, the particle free energy is higher in the right-handed state (DeltaG(t)=1.9(+/-0.I) kT), but it decreased (to 1.3(+/-0.1) kT) upon histone acetylation and the addition of phosphate, a potent tail destabilizer. Removal of the tails with trypsin further decreased DeltaG(t) (to 0.6 kT), and also induced a loss of supercoiling in both states, to DeltaLk(t)=-0.64(+/-0.03) and +0. 35(+/-0.05). The loop end-conditions, and hence the parameters of the DNA superhelix, were then calculated for both states using the explicit solutions to the equations of the mechanical equilibrium in the theory of elastic rod model for DNA. Whereas the pitch of the DNA superhelix may be approximately equal and opposite in the two conformations, its radius (r) was 20% larger in the right-handed conformation, confirming previous observations by electron microscopy of a tetrasome lateral opening in that conformation. The above supercoiling losses were found to reflect a further 3 % increase in r (to 23 %) upon removal of the tails in the right-handed conformation, and a 14 % increase in the left-handed conformation. The use of composite tetramers with one histone tail intact and the other removed showed these effects to be essentially due to the H3 tails. Altogether, these results show that the H3 tails oppose the tetrasome opening which is expected to be required to relieve the clash between the entering and exiting DNAs in the course of the transition, but which also appears to be intrinsic to the protein reorientation mechanism. We propose that the block against opening results from the H3 tails intercalating into the small groove of the double helix at +/-10 bp from the dyad, and acting as wedges against local DNA straightening. The tails (especially H3) may therefore regulate tetrasome chiral transition in vivo.

Acetylation↗

Clinicopathologic differentiation of atrophy of the pancreatic body and tail aplasia.

CONCLUSION: Congenital aplasia of the body and tail of the pancreas is derived from a defect of the dorsal pancreatic anlage and should not be considered a type of acquired atrophy of these structures. BACKGROUND: Congenital aplasia of the body and tail of the pancreas radiologically mimics acquired atrophy of the pancreatic body and tail. METHODS: Two patients with radiologically identified aplasia of the body and tail of the pancreas were studied clinicopathologically. RESULTS: An 82-yr-old man was diagnosed radiologically as having both carcinoma of the head of the pancreas and aplasia of the body and tail of the pancreas and underwent pancreatoduodenectomy. Pathologically the carcinoma was distributed in the anterosuperior part of the head of the pancreas, and spread into the duct of Santorini and intraductally to a portion of the main pancreatic duct beyond the junction of the ducts of Santorini and Wirsung. Consequently, obstructive pancreatitis of the body and tail of the pancreas developed, resulting in marked atrophy that mimicked aplasia of the body and tail of the pancreas. A 74-yr-old woman was diagnosed radiologically as having two carcinomas, one of the gallbladder and one of the stomach, and aplasia of the body and tail of the pancreas. During surgery, suspected parenchymal disappearance and fatty replacement in the body and tail of the pancreas were noted. Histologic examination of biopsy specimens from the body portion revealed atrophic pancreatic tissue surrounded by fat. Therefore, these patients had atrophy of the pancreatic body and tail.

Aged↗

Germination, Growth, and Nodulation of Sesbania rostrata Grown in Pb/Zn Mine Tailings

/ This study examined the possibility of growth, nodulation, and nitrogen accumulation of Sesbania rostrata in pure and amended Pb/Zn tailings. About 90% of seeds of S. rostrata germinated in pure Pb/Zn tailings, which contained high concentrations of Pb, Zn, Cu, and Cd. Although seedling growth suffered from the adverse environment of Pb/Zn tailings, they became established on tailings stands, in the greenhouse, as well as on the actual tailings dam, and completed their life cycle in 4 months. Dry matter production and nitrogen accumulation was 3200 kg/ha and 69.4 kg/ha, respectively in the actual tailings dam. Applying inorganic fertilizer to Pb/Zn tailings led to no obvious improvement in growth and nodulation of S. rostrata, while tailings amended by river sediment or domestic refuse rich in organic matter improved the growth and nodulation of the species. Azorhizobium caulinodans survived and formed N-fixing stem and root nodules in S. rostrata grown in pure Pb/Zn tailings with a nodule biomass exceeding 300 mg fresh matter per plant.KEY WORDS: Sesbania rostrata; Azorhizobium caulinodans; Pb/Zn mine tailings; Revegetation; Nitrogen fixation; Heavy metals; Tolerance

Journal Article↗

Facilitation of the tail-flick reflex by noxious cutaneous stimulation in the rat: antagonism by a substance P analogue.

Effects of noxious cutaneous stimulation on the tail flick reflex were examined in the anaesthetized rat. Noxious stimulation was applied by immersing the distal 4 cm of the tail in water at 55 degrees C for 1.5 min. The tail flick reflex was tested at 3 min intervals by applying a noxious radiant heat stimulus to a region of the tail 10 cm proximal to the tip. Tail immersion reduced reaction time to tail flick by 30% and 20% at 0.5 and 3.5 min after immersion, respectively. Reaction time returned to control at 6.5 min and tended to increase above baseline values at 9.5 and 12.5 min. Naloxone (10 mg/kg, i.p.) potentiated the effects of tail immersion on reaction time and prevented the increase above baseline. When the surface temperature of the skin used to evoke the tail flick reflex was raised by 10 degrees C using innocuous radiant heat, reaction time was not significantly different from the control, suggesting that an increase in skin temperature per se is insufficient to account for the response to immersion. Intrathecal administration of a substance P antagonist (1 nmol) attenuated the response to tail immersion. These results indicate that noxious cutaneous stimulation may release an agent in the spinal cord which facilitates the tail flick reflex, and that this agent is antagonized by a substance P antagonist.

Animals↗

Desipramine in small doses induces antinociception in the increasing temperature hot-plate test, but not in the tail-flick test.

The data reported for the antinociceptive effect of tricyclic antidepressants are conflicting. In this investigation, the effect of acute intraperitoneal (i.p.) administration of desipramine (2 and 5 mg/kg) was studied in rats, using the tail-flick test with simultaneous measurement of tail-skin temperature, and the increasing temperature hot-plate test. A constant negative correlation between tail-flick latency and tail-skin temperature, as described earlier, was also found in this study. Different ambient temperatures influenced the results of the tail-flick test. At an ambient temperature of 24-25 degrees C, desipramine gave rise to an apparent antinociception in the tail-flick test, which was found to be caused by a relative fall in tail-skin temperature. At 21-22 degrees C, no change in tail-flick latency was found after the administration of desipramine. In the increasing temperature hot-plate test, however, a dose-dependent antinociceptive effect of desipramine was observed at both ambient temperatures. The strong influence of moderate differences in ambient temperature on the results of the tail-flick test may explain some of the conflicting results reported in the literature. Whenever this test is used, the temperature of the tail should be recorded and taken into account in the evaluation of the data.

Animals↗

Energetics of DNA end binding by E.coli RecBC and RecBCD helicases indicate loop formation in the 3'-single-stranded DNA tail.

We examined the equilibrium binding of Escherichia coli RecBC and RecBCD helicases to duplex DNA ends possessing pre-existing single-stranded (ss) DNA ((dT)(n)) tails varying in length (n=0 to 20 nucleotides) in order to determine the contributions of both the 3' and 5' single strands to the energetics of complex formation. Protein binding was monitored by the fluorescence enhancement of a reference DNA labeled at its end with a Cy3 fluorophore. Binding to unlabeled DNA was examined by competition titrations with the Cy3-labeled reference DNA. The affinities of both RecBC and RecBCD increase as the 3'-(dT)(n) tail length increases from zero to six nucleotides, but then decrease dramatically as the 3'-(dT)(n) tail length increases from six to 20 nucleotides. Isothermal titration calorimetry experiments with RecBC show that the binding enthalpy is negative and increases in magnitude with increasing 3'-(dT)(n) tail length up to n=6 nucleotides, but remains constant for n > or =6. Hence, the decrease in binding affinity for 3'-(dT)(n) tail lengths with n > or =6 is due to an unfavorable entropic contribution. RecBC binds optimally to duplex DNA with (dT)6 tails on both the 3' and 5'-ends while RecBCD prefers duplex DNA with 3'-(dT)6 and 5'-(dT)10 tails. These data suggest that both RecBC and RecBCD helicases can destabilize or "melt out" six base-pairs upon binding to a blunt DNA duplex end in the absence of ATP. These results also provide the first evidence that a loop in the 3'-ssDNA tail can form upon binding of RecBC or RecBCD with DNA duplexes containing a pre-formed 3'-ssDNA tail with n > or =6 nucleotides. Such loops may be representative of those hypothesized to form upon interaction of a Chi site contained within the unwound 3' ss-DNA tail with the RecC subunit during DNA unwinding.

Base Sequence↗

SRY and human sex determination: the basic tail of the HMG box functions as a kinetic clamp to augment DNA bending.

Human testis-determining factor SRY contains a high-mobility-group (HMG) box, an alpha-helical DNA-binding domain that binds within an expanded minor groove to induce DNA bending. This motif is flanked on the C-terminal end by a basic tail, which functions both as a nuclear localization signal and accessory DNA-binding element. Whereas the HMG box is broadly conserved among otherwise unrelated transcription factors, tails differ in sequence and mode of DNA binding. Contrasting examples are provided by SRY and lymphoid enhancer factor 1 (LEF-1): whereas the SRY tail remains in the minor groove distal to the HMG box, the LEF-1 tail binds back across the center of the bent DNA site. The LEF-1 tail relieves electrostatic repulsion that would otherwise be incurred within the compressed major groove to enable sharp DNA bending with high affinity. Here, we demonstrate that the analogous SRY tail functions as a "kinetic clamp" to regulate the lifetime of the bent DNA complex. As in LEF-1, partial truncation of the distal SRY tail reduces specific DNA affinity and DNA bending, but these perturbations are modest: binding is reduced by only 1.8-fold, and bending by only 7-10 degrees . "Tailed" and truncated SRY complexes exhibit similar structures (as probed by NMR) and distributions of long-range conformational substates (as probed by time-resolved fluorescence resonance energy transfer). Surprisingly, however, the SRY tail retards dissociation of the protein-DNA complex by 20-fold. The marked and compensating changes in rates of association and dissociation observed on tail truncation, disproportionate to perturbations in affinity or structure, suggest that this accessory element functions as a kinetic clamp to regulate the lifetime of the SRY-DNA complex. We speculate that the kinetic stability of a bent DNA complex is critical to the assembly and maintenance of a sex-specific transcriptional pre-initiation complex.

Amino Acid Sequence↗