In order to determine the role of the clock SNP rs1801260 in circadian disruption, a study was performed in 85 overweight/obese Spanish women. After recruitment from a previous study, the participants had body measures taken, blood drawn for genotyping and took the Horne and Ostberg chronotype questionnaire. Additionally, circadian rhythms of body position, activity and wrist temperature were measured by a pair of sensors attached to the patients’ non-dominant arm.
The authors found that carriers of the C allele of rs1801260 had circadian rhythms with lowered amplitude, power and robustness compared to the TT geneotype. In general, C allele carriers had higher wrist temperature, especially during the early morning and late evening. In all of the measures of circadian activity, the C carriers had less daily activity and were phase delayed compared to TT controls.
rs1801260 in clock
NCBI Variation Viewer rs1801260
To discover the genetic causes of Familial Advanced Sleep Phase Syndrome (FASPS), the authors interviewed a family whose members suffer from FASPS. The authors found an A to G change in the csk1d gene that lead to a threonine to alanine mutation, by genetic screening of family members. The affected residue (amino acid 44) is conserved across species, and its mutation implies a role for csnk1d in the circadian clock.
The authors then assessed the function of the mutated protein by expressing it in bacteria. The mutant protein had a maximum reaction rate 60% that of the wildtype protein, as well as a 12% reduction in its Michaelis constant, as assessed by kinase assays.
The mutation was then introduced to fruit flies, and resulted in a slightly longer circadian period, as measured from periodogram analysis of locomotor activity.
Additionally, transgenic mice were generated via bacterial artificial chromosome cloning, and validated using RT-PCR with human primers. The authors found that the mice with the mutant csnk1d had a circadian period shorter by an estimated 24 minutes, and took longer to re-entrain to an LD 12:12 light cycle, compared to wild type mice.
The purpose of this study was to determine the genetic and biochemical basis for Familial Advanced Sleep Phase (FASPS) Syndrome. The authors mapped the gene to the qter region of Chromosome 2 via linkage analysis confirmed by PCR. One gene in this region is the human homolog of the Drosophila period gene (as confirmed by bacterial artificial cloning and mapping), mutations of which lead to abberant circadian rhythms.
An S662G mutation was indentified in this region via Single-strand conformation polymorphism. Many of the sampled individuals diagnosed with FASPS while none of the controls had this mutation.
This mutation was then characterized by several experiments. It was found that the mutation occured within the Casein Kinase 1 Epsilon (CSNK1E) binding domain by immunoprecipitation, and the mutated site is a phosphorylation target by electrophoresis. Additionally, the mutated site is C-terminal to many other phosphorylation sites by sequencing, and the authors hypothesize that it is the initiation site of a phosphorylation cascade. This hypothesis was confirmed by comparing a S662D mutant (similar to a phosphoserine) to the wildtype and S662G proteins.
To determine the effect of timing of eating on weight loss, a study of 420 overweight or obese patients recruited from Spanish nutrition clinics was performed. The authors found that those who ate lunch later tended to lose less weight than those who ate earlier, and carriers of the minor allele of the clock SNP rs4580794 ate their meals later than major allele homozygotes.
rs4580704 in clock
NCBI Variation Viewer rs4580704