The functional interplay between TNPO3, CPSF6 and HIV-1 CA

Lentiviruses can infect postmitotic cells, indicative of a role for the nucleocytoplasmic transport machinery. Genome-wide RNA interference screens identified transportin 3 (TNPO3) that may regulate human immunodeficiency virus type 1 (HIV-1) preintegration complex (PIC) nuclear import but plays no role during murine leukemia virus (MLV) infection. Independently, TNPO3 was shown to bind HIV-1 integrase (IN), a PIC component, suggesting a potential mechanism for nuclear import. We demonstrated direct binding between TNPO3 and several retroviral INs, which did not correlate with TNPO3 dependency profiles of the respective retroviruses. Infectivity assays employing HIV-1/MLV chimeric viruses ascertained that the capsid (CA) domain, but not IN, was the functional determinant of TNPO3 dependence. A carboxy-terminal truncation mutant of the serine-arginine rich (SR) protein family member, cleavage and polyadenylation specific factor 6 (CPSF6), CPSF6-358, which lacks its RS domain, was shown to restrict HIV-1 PIC nuclear import. We demonstrated that CPSF6 interacts with HIV-1 CA, and a single point mutation in CA, Asn74Asp (N74D), abolished this interaction. N74D also rendered HIV-1 TNPO3-independent and impaired cyclophilin A (CypA) binding to CA. The CA:CPSF6 binding interface, as described in a partial co-crystal structure, defined a surface pocket on CA that faces the CA hexamer:hexamer interspace. Infectivities and CA binding profiles of CA mutants within this pocket or with aberrant CypA-related phenotypes were assessed to compare their CPSF6-358 sensitivity and TNPO3 dependence, which largely correlated. We showed an overall correlation between the CPSF6/CPSF6-358 binding profiles of these HIV-1 CA mutants and their CPSF6-358 sensitivity, whereas TNPO3 binding and TNPO3 dependence did not correlate. Based on similar infectivity profiles of CA mutants and the loss of the RS domain from CPSF6-358 we tested for a direct interaction between CPSF6 and TNPO3. We demonstrated specific binding between recombinant TNPO3 and the CPSF6.RS domain. Mutagenesis experiments suggested a multicontact binding interface. The interaction was downmodulated by Ras-related nuclear protein (Ran)-GTP, indicating that CPSF6 is a bona fide import substrate of TNPO3. Our results support a model where TNPO3 regulates nuclear CPSF6 localization and that in its absence CPSF6 may restrict infection by directly interacting with HIV-1 CA at the hexamer:hexamer interface

Spinal fusion in facioscapulohumeral dystrophy for hyperlordosis A case report

Rationale: Facioscapulohumeral muscular dystrophy (FSHD) is the third most common muscular dystrophy, which is associated with facial, shoulder girdle, and paraspinal muscle atrophy. Most of the patients develop hypokyphosis and hyperlordosis in the course of the disease, to preserve standing posture. Corrective fusion is contraindicated in these patients as the surgery results with loss of compensatory hyperlordosis and leads to loss of trunk balance while standing. Although spinal fusion in neuromuscular scoliosis is a known treatment option, there are no studies in the literature on the spinal fusion of this specific patient group. Patient concerns: In this case report we have presented a 66-year-old woman, who was admitted with back and abdominal pain, inability to sit straight, abdominal discomfort, and numbness in the lower extremities after prolonged sitting. Diagnoses: The patient developed severe hyperlordosis causing intra-abdominal disorders, radicular symptoms, and sitting discomfort due to FSHD. Interventions: The patient underwent T2-S1 fusion and successful fusion was achieved. Outcomes: Individualized Neuromuscular Quality of Life Questionnaire (INQoL) was used to assess preoperative and 3 years postoperative functional outcomes. All domains and total score improved at the end of the follow-up period and successful fusion was verified radiologically. Lessons: This case suggests that spinal fusion may provide functional improvement in carefully selected patient groups. Patient stratification considering spinal disability is required for further studies in this specific indication

A MULTIDISCIPLINARY CLINICAL APPROACH TO FACIOSCAPULOHUMERAL MUSCULAR DYSTROPHY ORTHOPEDIC SURGERY IN FACIOSCAPULOHUMERAL DYSTROPHY

Background - Impaired shoulder function is the most disabling problem for daily life of Fascioscapulohumeral muscular dystrophy (FSHD) patients. Scapulothoracic arthrodesis can give a high impact to the functionality of patients. Here we report our experience with scapulothoracic arthrodesis and spinal stenosis surgery in FSHD patients

A novel shoulder disability staging system for scapulothoracic arthrodesis in patients with facioscapulohumeral dystrophy

Background: Scapulothoracic arthrodesis (STA) is a well-established surgical technique to provide scapular stabilisation in patients with facioscapulohumeral dystrophy (FSHD). There is no staging or scoring systems available to guide surgical decision. The aim of this study was to develop a staging system to evaluate the shoulder disability in patients with FSHD to guide surgical decision-making and assess its reliability among surgeons

The Requirement for Cellular Transportin 3 (TNPO3 or TRN-SR2) during Infection Maps to Human Immunodeficiency Virus Type 1 Capsid and Not Integrase ▿

Recent genome-wide screens have highlighted an important role for transportin 3 in human immunodeficiency virus type 1 (HIV-1) infection and preintegration complex (PIC) nuclear import. Moreover, HIV-1 integrase interacted with recombinant transportin 3 protein under conditions whereby Moloney murine leukemia virus (MLV) integrase failed to do so, suggesting that integrase-transportin 3 interactions might underscore active retroviral PIC nuclear import. Here we correlate infectivity defects in transportin 3 knockdown cells with in vitro protein binding affinities for an expanded set of retroviruses that include simian immunodeficiency virus (SIV), bovine immunodeficiency virus (BIV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), and Rous sarcoma virus (RSV) to critically address the role of integrase-transportin 3 interactions in viral infection. Lentiviruses, with the exception of FIV, display a requirement for transportin 3 in comparison to MLV and RSV, yielding an infection-based dependency ranking of SIV > HIV-1 > BIV and EIAV > MLV, RSV, and FIV. In vitro pulldown and surface plasmon resonance assays, in contrast, define a notably different integrase-transportin 3 binding hierarchy: FIV, HIV-1, and BIV > SIV and MLV > EIAV. Our results therefore fail to support a critical role for integrase binding in dictating transportin 3 dependency during retrovirus infection. In addition to integrase, capsid has been highlighted as a retroviral nuclear import determinant. Accordingly, MLV/HIV-1 chimera viruses pinpoint the genetic determinant of sensitization to transportin 3 knockdown to the HIV-1 capsid protein. We therefore conclude that capsid, not integrase, is the dominant viral factor that dictates transportin 3 dependency during HIV-1 infection

Identifying the Characteristics of Geriatric Patients who Referred to Outpatient Clinics of Physical Medicine and Rehabilitation: A Multicenter Descriptive Study

WOS: 000295573000008Aim: The aim of this study was to define the demographic and clinical characteristics of geriatric patients who referred to physical medicine and rehabilitation (PMR) outpatient clinics and to detect the differences between these characteristics in regard to age, sex and education level. Materials and Methods: 820 patients over 65 years old who attended 20 outpatient clinics were included in the study. In addition to demographic data, the complaints, comorbid diseases, pain levels, drugs being used, exercise and medical status of the patients were recorded. The effects of age, sex and education level on complaints, comorbid diseases and exercise habits were investigated. Results: The mean age of the patients was 71.7 +/- 5.5 years. 16.7% were living alone, 61.7% were housewives. 86% of the patients had one or more comorbid diseases - hypertension, gastric problems and heart disease were mostly encountered. The most common complaints were joint pain, fatigue and widespread body pain. The average number of pills taken per day was 4.02 +/- 0.9 (median 4), and the VAS pain score was 5.1 +/- 1.3 (median 5.0). History of falling was present in 16.5% of patients. 30.1% were routinely walking and 15.4% were performing exercise at home. In patients over 75 years, vertebral pain and deformity, urinary incontinence, eye problem, difficulty in swallowing, decrease in hearing, as well as balance and teeth problems were significantly more frequent than in younger subjects. Conclusion: Aged population constitutes most of the PMR outpatient clinic patients. Considering comorbid diseases, high number of daily taken drugs and falls, PMR specialist should be cautious in prescribing drugs and planning rehabilitation programme. For independence in activities of daily living in this age group, besides the musculoskeletal system, all other systems should be evaluated and a comprehensive geriatric rehabilitation programme should be constructed. Turk J Phys Med Rehab 2011;57:143-9