Alice Van den Broeke

Characterization of the ovine ribosomal protein SA gene and its pseudogenes.

BackgroundThe ribosomal protein SA (RPSA), previously named 37-kDa laminin receptor precursor/67-kDa laminin receptor (LRP/LR) is a multifunctional protein that plays a role in a number of pathological processes, such as cancer and prion diseases. In all investigated species, RPSA is a member of a multicopy gene family consisting of one full length functional gene and several pseudogenes. Therefore, for studies on RPSA related pathways/pathologies, it is important to characterize the whole family and to address the possible function of the other RPSA family members. The present work aims at deciphering the RPSA family in sheep.ResultsIn addition to the full length functional ovine RPSA gene, 11 other members of this multicopy gene family, all processed pseudogenes, were identified. Comparison between the RPSA transcript and these pseudogenes shows a large variety in sequence identities ranging from 99% to 74%. Only one of the 11 pseudogenes, i.e. RPSAP7, shares the same open reading frame (ORF) of 295 amino acids with the RPSA gene, differing in only one amino acid. All members of the RPSA family were annotated by comparative mapping and fluorescence in situ hybridization (FISH) localization. Transcription was investigated in the cerebrum, cerebellum, spleen, muscle, lymph node, duodenum and blood, and transcripts were detected for 6 of the 11 pseudogenes in some of these tissues.ConclusionsIn the present work we have characterized the ovine RPSA family. Our results have revealed the existence of 11 ovine RPSA pseudogenes and provide new data on their structure and sequence. Such information will facilitate molecular studies of the functional RPSA gene taking into account the existence of these pseudogenes in the design of experiments. It remains to be investigated if the transcribed members are functional as regulatory non-coding RNA or as functional proteins.

The effect of crude protein reduction on performance and nitrogen metabolism in piglets (four to nine weeks of age) fed two dietary lysine levels1

Lowering the CP level in piglet diets reduces the risk of post-weaning diarrhea and N excretion to the environment. The question remains at what point CP becomes limiting. An experiment was designed with 2 standardized ileal digestible (SID) Lys levels (10 and 11 g) and 6 CP levels (140, 150, 160, 170, 180, 190 g/kg) in a 2 x 6 factorial design (with 6 pens of 6 animals each per treatment). Linear and quadratic (QP) mixed models of performance in function of CP were fitted to study the effect of Lys and CP and their interaction. To determine optima, QP models and broken line models with linear (BLL) or quadratic (BLQ) ascending portions were fitted through the data. It was hypothesized 1) that the response to a decreasing digestible CP level could be described with broken line models and 2) that the breakpoint of these models is dependent on the dietary SID Lys level. Decreasing the CP level decreased ADG (P < 0.001). For G:F, the effect of decreasing CP level depended on the SID Lys level (P of the interaction = 0.028 in the linear model and P = 0.002 in the QP model). According to the BLL model, with 11 g SID Lys in the diet, G:F started to decline with CP levels below 176 g CP (SID Lys:apparent total tract digestible (ATTD) CP = 0.077), and with 10 g SID Lys, CP levels below 165 g/kg (SID Lys:ATTD CP = 0.075) depressed performance. Serum creatinine levels showed a linear decrease with increasing Lys:CP levels (P < 0.001). Across both SID Lys levels, when fitting a BLL model, minimal serum urea levels were reached at a Lys:CP ratio of 0.064. This seems to be the point where CP and not Lys limits muscle deposition. The small difference in breakpoint between serum urea level and performance suggests that the composition of nonessential AA may be also at stake. The effect of decreasing CP level depends on SID Lys and using a maximal SID Lys:CP ratio may be useful for optimizing the AA profile of dietary CP. When the Lys:CP ratio exceeds 0.064 (SID Lys:ATTD CP above 0.079), protein and not individual AA limits growth in most piglets between 4 and 9 weeks of age.